tmpF02E TMP

4 12
NF B is a centralHOME
regulatorQUESTIONS
of protein quality
JOBScontrol in responseSearch
to protein aggregation stresses via autophagy modulat... Add new
Article Apr 2016 Molecular biology of the cell
Download
Andre Patrick Arrigo 42.20

Emeritus professor
Emeritus professor Add a new Article

Centre Lon Brard, Lyon Apoptosis, Cancer and Develop...
Overview
NF
B isTimeline
Contributions
a central regulator
Info
of protein
Stats Scores
quality control in response to protein

aggregation stresses via autophagy modulation

Centre Lon Brard
What are you working on Apoptosis, Cancer and Development Laboratory
Mathieu Nivon*, Loc Fort*, Pascale Muller*, Emma Richet* ||

Lyon, France
, Stphanie Simon*,
right now? # @ ,~
Baptiste Guey* , Malenn Fournier*, Andr-Patrick Arrigo* , Claudio Hetz , Julie D.
Add your current project to share updates on your work
and get noticed for your research. Atkin and Carole Kretz-Remy*
About
I have studied Heat shock proteins (Hsps), particularly the small
ones (Hsp27, alphaB-crystallin) and participated in the discovery
* Universit de Lyon, F-69622, Lyon,
Enter a title for your project Add
France; Universit
of aLyon 1, Villeurbanne;
crucial proteolytic CNRS,
machine that we called the proteasome.
UMR5310, INSERM U1217, Institut NeuroMyoGne.

Biomedical Neuroscience Institute, ICBM, Faculty of Medicine, University of Chile, Santiago,
Connect with more colleagues
Show your career's best
Add your other social network services to Mnd more
Chile
Use your proMle overview page to present yourself and your research. Customizing your proMle is
colleagues and share your achievements and
the best way to show your peers what you've been working on, create exposure for your current updates.
projects, and start building ~Center
your network.for Geroscience, Brain Health and Metabolism (GERO), Santiago, Chile.
Connect now

Department of Biomedical Sciences, Faculty of Medicine & Health Sciences, 2 Technology
Place, Macquarie University, NSW, 2109, Australia.
191 6.16k 12,280 661.97 123 Skills and expertise (32)
Publications Reads Citations Impact Points Full-texts View stats
10 Cell Biology 8 Heat Shock Proteins 6 Apoptosis
Running title: NFB tunes protein quality control 6 Cancer Biology 6 Reactive Oxygen Species
Featured publications
Article: NF B is a central regulator of protein quality

control in response to protein aggregation stresses via Top co-authors View all
Corresponding
autophagy modulation author: Carole Kretz-Remy, INMG, CNRS UMR5310, INSERM U1217 Bt.
Carole Kretz 32.81 220.82 (43)
Mathieu Nivon Loc Fort Pascale Muller Emma Richet Claude Bernard University Lyon 1
G. Mendel,
Stphanie 16 Rue
Simon Baptiste Dubois,
Guey Malenn Universit
Fournier Andr- Claude Bernard Lyon 1, F-69622 Villeurbanne Cedex,
Patrick Arrigo Claudio Hetz Julie D Atkin Carole Kretz-Remy
Chantal
France. Mailing address: carole.kretz@univ-lyon1.fr; Phone Diaz-Latoud
number: +33 472 44 79 57; Fax
28.03 126.96 (32)
Source
63 number:
0 +33 472 43 26 85. Claude Bernard University Lyon 1
Reads Citations
Present addresses: Stphanie Simon 25.30 70.79 (19)
Universit Paris-Est Crteil Val de Marne ...

CRUK Beatson Institute for Cancer Research, College of Medical Veterinary and Life
Article: Meet Our Associate Editor:
Catherine Paul 30.45 151.49 (16)
Sciences,
Andre-Patrick ArrigoUniversity of Glasgow, Glasgow, UK Ecole Pratique des Hautes Etudes
||
OMNT CNRS/CEA UMS2920, Campus Minatec, 17 rue Florence
des Martyrs, F-38954 Grenoble
1 0 Manero 21.59 66.41 (13)
University of Angers
ReadCedex,
Citations
France

IMRB, INSERM, U955, Equipe 5, F-94000, Crteil, France
#
INSERM U1052, CNRS UMR5286, Centre de Recherche en author
Did this researcher Cancrologie
this publicationde
with Lyon,
you? F-
Chapter: Immense Cellular Implications Associated to
Small StressLyon,
69000 Proteins Expression: Impacts on Human
France
Pathologies Christiane Richter-Landsberg
@
Apoptose,
Andre Patrick Cancer
Arrigo Benjamin et Dveloppement,
Ducarouge Fabrice Lavial UMR INSERM 1052-CNRS
Chapter: The Role 5286, Centre
of Heat Shock Proteins de
during
Benjamin Gibert Neurodegeneration in Alzheimers, Parkinsons and
Recherche sur le Cancer de Lyon, Universit
H... Lyon1, France
Andreas Wyttenbach Andr Patrick Arrigo
45 1 Christiane Richter-Landsberg
Source
Reads Citation No preview Chapter May 2009
Skip No Yes
Questions Supplemental Material can be found at: 1

http://www.molbiolcell.org/content/suppl/2016/04/10/mbc.E15-12-0835v1.DC1.html
Ask and answer questions to connect with other researchers in your Meld.
149 Following View all
Topics (7)
Molecular Biology Cell Biology Cancer Research Methods Cancer Biology
Page 1
4 12
HOME QUESTIONS JOBS Add new

ABSTRACT
Emeritus professor
During
Emeritus cell
professor Addmutations
life, proteins often misfold, depending on particular a new or Article
environmental
changes, which may lead to protein aggregates that are toxic for the cell. Thus protein
aggregates are the root cause of numerous diseases called protein conformational
Overview diseasesTimeline
Contributions such as myofibrillar
Info
myopathy
Stats
Scoresor familial amyotrophic lateral sclerosis. To fight

against aggregates, cells are equipped with protein quality control mechanisms. Here, we
report that NFB transcription factor is activated by misincorporation of amino acid analogs
Centre Lon Brard
What are you working
into proteins, on of proteasomal activity, expression Apoptosis,
inhibition
Cancer and Development Laboratory
of the R120G mutated form of
Lyon, France
right now?HspB5 associated to myofibrillar myopathy- or expression of the G985R and G93A mutated
forms
Add your current project of superoxide
to share dismutase
updates on your work 1 linked to familial amyotrophic lateral sclerosis. This non-
and get noticed for your research. About
canonical stimulation of NFB triggers the upregulation of BAG3 and HspB8 expression, two
activators of selective autophagy, which relocalizeones
to (Hsp27,
protein aggregates.
alphaB-crystallin) Then, inNFB-
and participated the discovery
Add
of a crucial proteolytic machine that we called the proteasome.
dependent autophagy allows to clear protein aggregates. Thus, NFB appears as a central
and major regulator of protein aggregates clearance, by modulating autophagic activity. In
this context, the pharmacological stimulation of this quality
Connectcontrol pathway
with more might represent a
colleagues
valuable strategy for therapies against protein conformational diseases.
projects, and start building your network.
INTRODUCTION Connect now
Proteins fold into specific three-dimensional structures to acquire their functional native
states. However, proteins can misfold as a result of inefficient protein biogenesis, mutations
191 6.16k 12,280 661.97 123
Skills and expertise (32)
Publications Reads
or inefficient
Citations
translocated
Impact Points
secretory
Full-texts
precursors
View stats
(Herczenik and Gebbink, 2008). In addition,
protein misfolding can be promoted by environmental stresses such as elevated
temperatures, exposure to chemicals or heavy metals,6 bacterial/viral
Cancer Biology 6 Reactive Oxygen Species
infection, production of
reactive oxygen species as well as by physiological changes such as aging (Haigis and
Article: NF B is a2010;
Yankner, regulatoretofal.,
central Chen protein qualityBecause of their altered spatial arrangement, misfolded
2011).
autophagy
proteins modulation
are non functional (loss of function effect) and tend to exhibit their hydrophobic
Carole Kretz 32.81 220.82 (43)
domains,
Stphanie driving
Simon Baptiste Gueyinappropriate
Malenn Fournier proteins
Andr- association and protein aggregation (Herczenik and
Gebbink, 2008); moreover, because protein aggregates Chantal
can Diaz-Latoud
trap other
functional cellular
28.03 126.96 (32)
Source
63 components,
0 Claude Bernard
they are often toxic for the cell (gain of function University
effect) Lyon 1
as evidenced by the
Reads Citations
growing list of protein conformational diseases, which result from
Stphanie the
Simon cellular accumulation
25.30 70.79 (19)
of proteotoxic misfolded/aggregated proteins (Hightower, 1991; Holmes et al., 2014).
Consequently,
Andre-Patrick Arrigo cells can detect the presence of abnormal proteins
Ecole Pratique des(for pioneer
Hautes Etudes study see
(Edington et al., 1989)) and have developed machineryFlorence
that Manero
maintains the health of its
1 0 21.59 66.41 (13)
Readproteome
Citations by: refolding, sequestering or degrading misfolded proteins. Refolding is achieved
by molecular chaperones (Vabulas et al., 2010); spatial sequestering is for example fulfilled
by addressing aggregates to the microtubule-organizing center (MTOC), forming a unique
Did this researcher author this publication with you?
Small Stress Proteins Expression: Impacts on Human
Andre Patrick Arrigo Benjamin Ducarouge Fabrice Lavial Chapter: The Role of Heat Shock Proteins during
Benjamin Gibert Neurodegeneration in Alzheimers, Parkinsons
2 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 2
4 12

large storage-bin structure called aggresome (Garcia-Mata et al., 1999). At last,
Emeritus professor
degradation
Emeritus professor Add a new
is principally accomplished by the ubiquitin proteasome Article
system (UPS), which
clears most soluble misfolded proteins (Goldberg, 2003), and the lysosomal compartments
such as the autophagy-lysosomal pathway.
Overview Macroautophagy
Contributions Timeline (hereafter

Info referred
Stats
toScores
as autophagy) is a self-eating process starting with

the formation of double-membrane autophagosomes that engulf parts of the cytoplasm and
fuse with acidic lysosomes to form autolysosomes (Wirth etCentre
al., Lon
2013; Wilson et al., 2014).
Brard
What are you
Then,working onmaterial is degraded by acid hydrolases
the enclosed Apoptosis, Cancer and Development Laboratory
and released for recycling.
Lyon, France
right now?Autophagy occurs at basal level in most tissues and allows the degradation of long-lived
proteins
Add your current project to shareand cytoplasmic
updates on your work organelles, contributing to the routine turnover of cellular trash
(Mehrpour et al., 2010). Under nutrient starvation, autophagy
and get noticed for your research. About is upregulated as an energy
salvage process (Kuma et al., 2004), but it can also be activated under non-metabolic
Add
stresses such as accumulation of aggregated proteins.of aOf
crucial proteolytic machine that we called the proteasome.
interest, loss of autophagy function
may cause an accumulation of misfolded/aggregated proteins (Rubinsztein, 2006; Williams et
al., 2006; Villella et al., 2013). While autophagy Connect
has long been described as a bulk
with more colleagues
degradation process, it is now largely accepted that autophagyAddcan be selective
your other social networkvia thetouse
services Mnd more
of autophagic
the best way to show your peers receptors
what you've been working on, createand adaptors
exposure (such
for your current as p62 or BAG3), linking cargos to the
updates.
projects, and start building autophagosomal
your network. membrane protein LC3-II (Behrends and Fulda, 2012). Recently, we
Connect now
showed that selective autophagy is activated by protein aggregates generated by
hyperthermia, via the stimulation of NFB (Nivon et al., 2009; Nivon et al., 2012).
191 6.16k
NFB 12,280
is a key 661.97
transcription123
factor of the cellular stress
response. This inducible factor is
10 Cell Biology
inactivated by its binding to members of the IB family (Hayden 8and
Heat Shock Proteins
Ghosh, 2004); 6 Apoptosis
hence,
6 Cancer Biology 6 Reactive Oxygen Species
activation of NFB requires its dissociation from IB subunits. This event mostly occurs
through the canonical pathway consisting of IKK phosphorylation, which in turn leads to
phosphorylation, ubiquitination and degradation of IB by the 26S proteasome (Hayden and
autophagy
Ghosh, modulation
2004). However, some inducers stimulate NFB through non-canonical pathways.
Carole Kretz 32.81 220.82 (43)
Stphanie
Indeed,Simon UV
Baptiste
orGueyamino
Malenn Fournier
acid analog
Andr- treatments induce IB degradation without prior
phosphorylation (Kretz-Remy et al., 1998; Li and Karin, 1998);
Chantal Diaz-Latoud
we also demonstrated that
28.03 126.96 (32)
Source
63 0 Claude Bernard University Lyon 1
heat stress induces NFB/IB dissociation independently of IB phosphorylation or
Reads Citations
degradation (Nivon et al., 2009). Stphanie Simon 25.30 70.79 (19)
Here,
Article: we
Meet Ouraimed atEditor:
Associate determining if NFB could act as a central modulator of protein quality
Andre-Patrick Arrigo
control (PQC) upon proteotoxic stress. We thus tested Ecole
NFPratique des Hautes Etudes
B involvement in PQC after
1 various
0 Florence
protein aggregation stresses such as amino acid Manero treatment,
analog 21.59 66.41proteasome
(13)
Read Citations
inhibition by MG132 or overexpression of mutated forms of proteins prone to form
aggregates. The amino acid analogs canavanine and azetidine mimic natural arginine and
proline
Chapter: respectively;
Immense they can
Cellular Implications escapetodetection
Associated by theDidprotein
this researcher author thismachinery
synthesis publication withand
you? thus
3 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 3
4 12

are incorporated into nascent polypeptides promoting translational errors and irreversible
Emeritus professor
protein
Emeritus misfolding,
professor Add
resulting in protein aggregation (Hightower, a new Article
1980Hightower, 1980 #145;
Kretz-Remy et al., 1998; Rodgers and Shiozawa, 2008). MG132 reversibly inhibits the
chymotrypsin and caspase-like activities of the 26S proteasome and thus, increases the level
Overview of poly-ubiquitinated
Contributions Timeline proteinsStats
Info and
induces
Scores protein aggregates and aggresome formation
(Shen et al., 2011). Interestingly, cardiac dysfunctions, cataract formation but also
neurodegenerative diseases (Alzheimers, Parkinsons, Amyotrophic
Centre Lon Brard
Lateral sclerosis,
Huntingtons) on
are associated Apoptosis, Cancer and Development Laboratory
with decreased proteasome activity (Dahlmann, 2007). HspB5
Lyon, France
right now?(CRYAB gene) is a member of the small heat shock protein family. It is abundantly expressed
in lens,
Add your current project cardiac
to share updatesand skeletal
on your work muscles and, by chaperoning desmin and actin, participates in
maintaining cytoskeletal integrity (Simon et al., 2013).
and get noticed for your research. AboutThe R120G missense mutation of
HspB5 was the first to be identified in French human patients and causes cataract,
Add
cardiomyopathy and myofibrillar myopathy (Vicart etof aal.,
crucial proteolytic machine that we called the proteasome.
1998). At the cellular level, this
mutation promotes HspB5 aggregation with intermediate filaments (desmin, vimentin) that
ultimately lead to aggresome formation (Simon et al., Connect
2007b;with
Elliott et al., 2013). Cytoplasmic
more colleagues
Cu, Zn-superoxide dismutase (SOD1) is an enzyme responsible
Addfor
yourscavenging free
other social network radicals.
services to Mnd more
More
the best way to show your peers than
what you've140
been point
working mutations found
on, create exposure in current
for your the SOD1 peptide sequence are associated with 20%
updates.
projects, and start building of
your network.
familial amyotrophic lateral sclerosis (fALS) cases (Rosen et al., 1993). ALS is a
Connect now
progressive and fatal adult-onset motor neuron disease characterized by premature loss of
spinal, cranial and cortical neurons, muscle weakness, atrophy and paralysis. At the cellular
191 6.16k
level, 12,280 661.97 mutations
most of SOD1 123 alter its tertiary structure, leading (32)
Skills and expertise to protein aggregation
(Turner et al., 2005; Kim et al., 2014). 10 Cell Biology 8 Heat Shock Proteins 6 Apoptosis
In this study, we show that NFB-depleted cells are unable

6 Cancer Biology to6 induce autophagy
Reactive Oxygen Species and
Featured publications accumulate more aggregates than control cells in response to a wide range of protein
aggregation
Article: stresses.
NF B is a central regulatorOur dataquality
of protein indicate that, in addition to heat stress, at least four other
different
autophagy protein aggregation stresses can activate NF
modulation B transcription factor through a non-
Carole Kretz 32.81 220.82 (43)
Mathieu Nivon Loc Fort Pascale Muller Emma Richet
canonical pathway that, in turn, induces
Stphanie Simon Baptiste Guey Malenn Fournier Andr-
the autophagic process. Moreover,
Claude Bernard we1 demonstrate
University Lyon

that upon those stresses, NFB activation enhances theChantal
expression
Diaz-Latoudof
BAG3 and HspB8
28.03 126.96 (32)
Source
63 proteins,
0 which relocalize to a vast majority of protein aggregates and activate
Claude Bernard University Lyon 1 selective
Reads Citations
autophagy and protein aggregates clearance. Thus NFB can be
Stphanie considered
Simon as
25.30 70.79 a central
(19)
regulator of PQC via regulation of autophagy in response to protein aggregation stresses.
Andre-Patrick Arrigo Ecole Pratique des Hautes Etudes
1 0 Florence Manero 21.59 66.41 (13)

Read Citations
Chapter: Immense Cellular Implications Associated to Did this researcher author this publication with you?
4 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 4
4 12

RESULTS
Emeritus professor
Amino
Emeritus acid
professor analogs and MG132 treatments, expression ofAdd a newforms
mutated Article
of HspB5 or
SOD1 induce the formation of protein aggregates.
We previously reported that heat stress activates NFB transcription factor via the induction
Overview
of proteinTimeline
Contributions aggregation
Info(Kretz-Remy
Stats

etScores
al., 2001). We therefore investigated whether other
types of protein aggregation stresses could activate NFB. We first analyzed, by
immunofluorescence (Figure1, A to E) and filter trap assayCentre
(supplementary
Lon Brard figure S1), the
protein onstatus of HeLa cells upon four differentApoptosis,
aggregation
Cancer and Development Laboratory
protein aggregation stresses:
Lyon, France
right now?amino acid analogs and MG132 treatments or expression of the mutated forms of HspB5 and
SOD1,
Add your current project which
to share areonprone
updates to
your work form aggregates. The distribution of multi-ubiquitinated proteins
and get noticed for your research.
was rather diffused in the cytoplasm in non-treated About
cells (Figure 1, A to C, NT panels). In
contrast, 5mM canavanine or azetidine treatments induced the
ones (Hsp27, formationand
alphaB-crystallin) ofparticipated
small cytosolic
in the discovery
Add
granules containing multi-ubiquitinated proteins in around 20% of the cells (Figure 1, A and B
graphs). Moreover, 15 mM of the same compounds induced, in a higher percentage of cells
(33 to 38%), aggregates of larger size, co-localizedConnect
with pericentrin, a component of the
MTOC, and sometimes gathered around the nucleus, suggesting aggresome
Add your other formation
social network services to Mnd more
(Figure
the best way to show your peers 1, Abeen
what you've and B).on,Filter
working trap analysis
create exposure confirmed
for your current the accumulation
updates. of protein aggregates
with more than 4 fold increase in the level of multi-ubiquitinated protein aggregates after
Connect now
amino acid analog treatments in comparison to non-treated cells (supplementary figure S1,
A). Similarly, we detected that increasing concentrations of MG132 induced the formation of
191 6.16k 12,280 positive
multi-ubiquitin 661.97 inclusions
123 of larger
size and Skills and expertise
increased (32)
frequency in the perinuclear
region (aggresome-like structures) of treated cells (Figure 1C, pictures and graph). Indeed,
38% of 2.5 M MG132-treated cells contained aggregates, with
6 Cancer 10%6ofReactive
Biology these cells
Oxygen containing
Species
Featured publications aggregates larger than 3 m. In comparison 74% of 5 M MG132-treated cells contained
Article: NF B is a central
aggregates, withregulator
43% ofofthese
proteincells
qualitycontaining aggregates with a size superior to 3 m. Filter
trap assay
autophagy indicated at least 8 fold increased level of multi-ubiquitinated protein
modulation aggregates
Carole Kretz 32.81 220.82 (43)
after 2.5 or 5 M MG132 treatments
(Figure S1, B). AsClaude
for Bernard
overexpression
University Lyon 1 experiments,

pcDNA3-HspB5WT transfected cells (HspB5wt) showed a Diaz-Latoud
Chantal homogeneous
and diffuse
28.03 126.96 (32)
Source
63 localization
0 of HspB5 (Figure 1D), as previously describedClaude
(Simon etUniversity
Bernard al., 2007a).
Lyon 1 In contrast,
Reads Citations
when cells expressed the missense-mutated form of HspB5 (HspB5R120G,
Stphanie Simon 25.30 found in human
70.79 (19)
disease), the protein was localized in cytoplasmic aggregates in 40% of the cells, with 14% of
the aggregates larger than 3 m. Moreover, some of theCatherine
Paul 30.45 151.49 (16)
aggregates were gathered in the
Ecole Pratique des Hautes Etudes
perinuclear region (co-localization with pericentrin). Filter trap data indicated around an 8 fold
1 0 Florence Manero 21.59 66.41 (13)
University
toofcells
Angers expressing wild type
Readincrease
Citationsof HspB5 aggregates in those cells in comparison
HspB5 (Figure S1, C). Similar observations were made with SOD1 protein, when expressed
in HeLa cells. The localization of wild type SOD1-EGFP was diffuse in the cytoplasm and the
nucleus
Small (Figure
Stress Proteins 1E). In contrast,
Expression: the expression
Impacts on Human of the G93A mutated form of SOD1 induced
5 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 5
4 12

the appearance of cytoplasmic or juxta-nuclear EGFP inclusions in 17% of the cells.
Emeritus professor
SOD1G85R
Emeritus professor Add a
expression induced similar inclusions with a higher new Article
frequency of appearance
(27%) but with a similar percentage of cells containing aggregates with a size superior to 3
m (around 9%). Filter trap analysis indicated that SOD1G85R and SOD1G93A proteins
Overview increasedTimeline
Contributions the level ofInfo
proteinStats
aggregates respectively by 8 and 4 fold compared to wild type
Scores
SOD1 (Figure S1D). Hence, by submitting HeLa cells to those diverse stresses, we were
able to control protein misfolding and to induce various typesCentre
(quantitatively
Lon Brard
and qualitatively)
of protein onin the cells.
aggregates Apoptosis, Cancer and Development Laboratory
Lyon, France
right now?
Amino
Add your current project acid
to share analogs
updates on yourand
work MG132 treatments or expression of HspB5 and SOD1 mutants
activate NF
B through a non-canonical pathway. About
We then tested if protein aggregation stresses could activate NFB
ones (Hsp27, transcription
alphaB-crystallin) factor,in as
and participated we
the discovery
Add
previously reported for heat and amino acid analog treatments (Kretz-Remy et al., 1998;
Kretz-Remy et al., 2001). NFB activity was quantified in HeLa cells constitutively expressing
pNFBluc reporter vector (see material and methods) and
Connect with moresubmitted
colleagues to increasing
Add yourwith
concentrations of azetidine, canavanine and MG132 or transfected other social
wild network services
type or to Mnd more
mutated
the best way to show your peers
HspB5 what you've
andbeen working on, create exposure
SOD1-expression for your current
vectors (Figure 2). B-dependent
updates.luciferase reporter gene
revealed that, increasing concentrations of azetidine or canavanineConnect
gradually
now stimulated
NFB activity by around 2.5 fold for azetinine and 5 fold for canavanine treatments (Figure
2A) as we previously showed (Kretz-Remy et al., 1998). Noteworthy, this activation was not
Publications as
Reads intense
Citationsas that observed
Impact Points after a 43C
Full-texts heat shock treatment (around 24 fold). Similarly,
View stats
increasing concentrations of MG132 gradually activated NFB activity up to 6 fold (Figure
2B). Moreover, the overexpression of the mutated-form of HspB5 stimulated NFB activity by
around 2 fold whereas overexpression of wild type HspB5 did not. Indeed, NFB activity in
HspB5wt-expressing
control cells was
in response to protein aggregation similar
stresses via to the one observed in untreated or empty vector-View all
Top co-authors
autophagy modulation
transfected (Cont) cells (Figure 2C). The equivalent expression
Carole Kretz levels of wild
32.81 220.82 (43) type and
mutated HspB5 were established by western bot analysis (Figure 2C insert). Similarly,
Chantal Diaz-Latoud
equivalent expression of G93A and G85R mutated forms of SOD1
28.03 increased
126.96 (32) NFB activity
Source
by
Reads around
Citations2 fold whereas wild type form did not (Figure 2D and Figure 2D insert). We then
Stphanie Simon 25.30 70.79 (19)
performed the same experiment in cells expressing either Universit
a dominant Paris-Estnegative
Crteil Val demutant
Marne ... of the
Article:
IKK Meet Our Associate
subunit of IKKEditor:
(pRK5-IKK(K44A)) or a dominant Catherine
negative mutant of IB (pLXSN
Paul 30.45 151.49 (16)
IBM), which both abrogate IB phosphorylation and thus the canonical activation
1 pathway
0 Florence Manero 21.59 66.41 (13)
of NFB (Figure 2E). NFB stimulation by proteinUniversity
aggregation
of Angers stresses was similar
Read Citations
in presence and absence of those dominant negative mutants. In contrast, NFB induction by
TNF treatment, which depends on the canonical pathway, was strongly impaired by the
presence
Small of those
Stress Proteins dominant
Expression: negative
Impacts on Human mutants (Figure 2E). Thus, NFB activation by those
6 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 6
4 12

protein aggregation stresses seems independent of IB phosphorylation/degradation.
Emeritus professor
Taken
Emeritus together,
professor Add a newstresses
our results demonstrate that various protein aggregation Articleare able to
activate NFB, all via a non-canonical pathway. Moreover, they suggest that NFB is a
central sensor of cellular protein aggregation stress.
Overview
Contributions Timeline
Info
Stats
Scores
Amino acid analogs, MG132 treatments and expression of mutated forms of HspB5 or
SOD1 activate autophagy flux in an NF
B-dependent manner.
Centre Lon Brard
What are you workingreported
We previously on that protein aggregation stress induced
Apoptosis, Cancer and Development Laboratory
by heat shock activates
Lyon, France
right now?NFB-dependent autophagy. We thus wondered whether the aggregation stresses studied
Add your current project
hereto could
share updates
induceon your
the work
same process. To this end, HeLa cells were submitted to increasing
concentrations of azetidine, canavanine or MG132 and LC3-II level was quantified by
western blot as a read-out of the abundance
Add of autophagosomes in cells. and
ones (Hsp27, alphaB-crystallin) We observed
participated in thethat
discovery
LC3-II level was augmented in cells treated with increasing concentrations of amino acid
analogs (Figure 3A) or MG132 (Figure 3B) compared to non-treated cells. Regarding
overexpression of HspB5, transfection by control plasmid or more
Connect with overexpression
colleagues of wild type
Add your other social network
HspB5 did nearly not modify LC3-II levels in comparison to non-transfected cellsservices
(1.2 toratio
Mnd more
versus
the best way to show your peers 1 forbeennon-transfected
what you've cells).
working on, create exposure Incurrent
for your contrast, overexpression of HspB5R120G doubled
updates.
LC3-II level (Figure 3C). The same results were obtained with G93A and now
Connect G85R mutated
forms of SOD1 whose overexpression increased by around 2 fold LC3-II levels, whereas wild
type SOD1 did not (Figure 3D). Those results were confirmed by immunofluorescence
191 6.16k 12,280 661.97 123
Publications Reads
analysis in cellsImpact
Citations
expressing
Points
GFP-LC3 View
Full-texts
or stats
labeled with LC3 antibody (supplementary figure
S2). Indeed, whereas we observed a diffuse fluorescence in non treated cells or in cells
expressing wild type HspB5 or SOD1 (Figure S2 A6 toCancer
C, Biology 6 Reactive Oxygen Species
NT, Cont and wt panels), we
observed the formation of numerous fluorescent puncta in cells treated by amino acid
Article: NF B isand
analogs a central
by regulator
MG132of(Figure
protein quality
S2A) or in cells overexpressing HspB5R120G (Figure S2B)
or SOD1G93A-EGFP and SOD1G85R-EGFP (Figure S2C). An increased number of
Carole Kretz 32.81 220.82 (43)
Mathieu Nivon Loc Fort Pascale Muller Emma Richet Claude Bernardautophagic
University Lyon 1 flux
autophagosomes
Stphanie Simon Baptiste Gueycan be Fournier
Malenn either Andr-
the consequence of an increased or of an
inhibition of the maturation/degradation of autophagic vesicles.
Chantal To distinguish
Diaz-Latoud between those
28.03 126.96 (32)
Source
63 two events,
0 HeLa cells were stressed as described above Claude
in theBernard Universityof
presence Lyon
5 1M E64d and
Reads Citations
pepstatin A, two inhibitors of lysosomal cathepsins that abrogate autophagosome
Stphanie Simon 25.30 70.79 maturation
(19)
into autolysosomes (Figure 3, E to G). Upon stimulation of protein aggregation, addition of
these inhibitors
Andre-Patrick Arrigo further enhanced the already elevated level ofPratique
Ecole LC3-II des(see
Hauteswestern
Etudes blots and
quantification graphs), indicating that treatments by amino acid analog or MG132 and
1 0 Florence Manero 21.59 66.41 (13)
Readoverexpression
Citations of HspB5R120G, SOD1G85R or SOD1G93A activated the autophagic flux.
We next determined whether this increased autophagic flux upon protein aggregation
stresses was NFB-dependent, by quantifying autophagic
Did thisactivity
researcherin control
author HeLa with
this publication cells and in
you?
7 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 7
4 12

HeLa cells depleted from the p65 subunit of NFB (p65-KD#2). We observed that in non
Emeritus professor
treated
Emeritus p65-KD#2
professor Add a
cells, LC3-II level was very slightly increased, asnew Article
already observed in a
previous study in which we demonstrated a minor role of NFB in autophagosome maturation
(Nivon et al., 2012). However, those cells were shown to still be able to induce autophagy, for
Overview
instance Timeline
Contributions after TNF Info
treatment
Stats
et al.,
(NivonScores
2009). After azetidine, canavanine (Figure 4A)

and MG132 treatments (Figure 4B), we observed that LC3-II level was increased by about 3
fold in HeLa cells, whereas it was not in p65-depleted cells (p65-KD#2). Similarly, expression
Centre Lon Brard
What are you working(Figure
of HspB5R120G on 4C) or of SOD1G85R and SOD1G93A Apoptosis, Cancer and Development Laboratory
(Figure 4D) induced a 2 to
Lyon, France
right now?2.5 fold increase of LC3-II levels in control HeLa cells but not in p65-KD#2 cells submitted to
Add your current project
the tosame
share updates on your work
treatments. Therefore, our results indicate that NFB is a major activator of
autophagic flux in response to diverse protein aggregation stresses.
Add
NF
B-dependent autophagy is involved in the clearance of protein aggregates
generated by treatments with amino acid analogs or MG132 or by expression of HspB5
and SOD1 mutants Connect with more colleagues
Autophagy is a and
process involved in PQC viaisdegradation Add your
of protein other social network
aggregates serviceset
(Sridhar al.,more
to Mnd
Use your proMle overview page to present yourself your research. Customizing your proMle
the best way to show your peers what you've been working on, create exposure for your current
2012). We previously reported that NFB-dependent autophagyupdates.
helps the cell to recover from
thermal injury by clearing protein aggregates. We therefore analyzed the protein
Connect now aggregation
status in control and p65-depleted HeLa cells submitted to various protein aggregation
191 6.16k
stresses (Figure 5). Cell lines were submitted to amino acid analog treatments and protein
12,280 661.97 123 Skills and expertise (32)
Publications aggregation
Reads Citations was monitored
Impact Points by immunofluorescence
Full-texts View stats and analysis of multi-ubiquitinated
proteins labeling (Figure 5A). In untreated control and p65-depleted HeLa cells (NT), labeling
of multi-ubiquitinated proteins was faint and rather diffuse into the cytoplasm, even if some
rare and small cytoplasmic inclusions could be detected in p65-deficient cells. This is
consistent
control in responsewith the aggregation
to protein fact that stresses
NFB via has a minor role in basal protein quality control by
Top co-authors View all
220.82 et
(43)al., 2012).
modulating autophagosomes maturation as we previously Carole
demonstrated
Kretz 32.81 (Nivon
Stphanie
AfterSimon
15 mM Baptiste Guey Malenn
azetidine or Fournier Andr- treatments,
canavanine we could distinguish some small inclusions
Chantal Diaz-Latoud
in respectively 31% and 38 % of control HeLa cells with rare (1.8% and 3.8%) aggregates of
28.03 126.96 (32)
Source
63
larger0 size (graph Figure 5A). In contrast, in p65-depleted cells, those inclusions were more
Claude Bernard University Lyon 1
Reads Citations
abundant and of larger size. Indeed 58 to 62% of the Stphanie Simon 25.30 aggregates,
cells contained 70.79 (19) with
aggregates
Article: superior
Meet Our Associate to 3
Editor: m in 7% of the azetidine-treated and in 14% of the canavanine-
Andre-Patrick
treatedArrigo
cells. Similar observations were performed in cells Ecole
treated with
Pratique des MG132 (Figure 5B), in
Hautes Etudes
1 which
0 we distinguished inclusions in 38% and 70% of control HeLa
Florence cells
Manero treated
21.59 66.41with
(13) 2.5 M
Read
or 5Citations
M MG132 respectively. However, this proportion of aggregates-containing cells
increased to 70% (2.5 M) or 98 % (5 M) in NFB-deficient cells, with also an increased
proportion
Chapter: ImmenseofCellular
aggregates of larger
Implications sizes.
Associated to Those large Did
inclusions co-localized
this researcher with with
author this publication pericentrin,
you?
8 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 8
4 12

indicating the formation of an aggresome-like structure. After cell transfection with pcDNA3-
Emeritus professor
BCWT
Emeritus professor(HspB5wt), Addinto
we observed a diffuse distribution of HspB5 a new Article with the
the cytoplasm
appearance of more dense structures only in 7.5% of p65-depleted cells. Interestingly,
expression of the R120G mutated form of HspB5 induced the formation of small inclusions
Overview
only
Contributions in 41% of the control
Timeline Info
HeLa
Statscells with
Scoresonly 6.5% of the cells containing large inclusions.

In contrast, this mutated protein induced the formation of aggresome-like structures in nearly
every p65-deficient cells with 50% of the cells containing large
Centresize inclusions (Figure 5C).
Lon Brard
Finally, on of SOD1G93A and SOD1G85R induced
the expression
mostly the formation of small
Lyon, France
right now?cytoplasmic inclusions in respectively 15% or 25% of control HeLa cells (Figure 5D).
However,
Add your current project inclusions
to share updates on yourwere
work detected in a majority, 45% and 70%, of p65-deficient cells
expressing SOD1G93A and SOD1G85R rather than the diffuse and widespread fluorescence
observed in control or p65-deficient cells expressing ones
SOD1wt. In addition,and
(Hsp27, alphaB-crystallin) theparticipated
percentage of
in the discovery
Add
cells containing large size inclusions was increased by approximately 3 fold in p65-KD#2 cell
line. Taken together our observations indicate that NFB-deficient cells have an impaired
autophagy induction in response to aggregation stresses, coinciding
Connect with an altered PQC and
clearance of protein aggregates. Add your other social network services to Mnd more
NF
B modulates BAG3 and HspB8 expression in responseConnect
to various
now
protein
aggregation stresses
Different studies have described the role of the BAG3-HspB8 complex in the selective
191 6.16k
removal
12,280
of misfolded
661.97
proteins by autophagy
123
(Carra etSkills

al., and expertise
2008a; (32) et al., 2008b; Carra,
Carra
2009). Moreover, we demonstrated in a previous work that upon heat shock, NFB
modulates BAG3 and HspB8 levels (Nivon et al., 2012).
We thus wondered if other types of
protein aggregation stresses could act in the same way. To that extent, we quantified the
Article:
level NF of
B isBAG3
a centraland
regulator of protein
HspB8 quality in control and p65-depleted HeLa cells submitted to
proteins
azetidine, canavanine and MG132 treatments (Figure 6A). We observed an in-between 1.6 to
Carole Kretz 32.81 220.82 (43)
3 fold
Stphanie Simonincrease of BAG3
Baptiste Guey and HspB8
Malenn Fournier Andr- levels (graph FigureClaude
6A)Bernard University Lyon
in control cells1 treated with

amino acid analogs or proteasome inhibitor; this increase was
Chantal strongly
Diaz-Latoud reduced in p65-
28.03 126.96 (32)
Source
63 depleted
0 Claude Bernard
cells. Of interest, the expression of HspB5R120G mutant University
or Lyon 1
SOD1G85R and
Reads Citations
SOD1G93A also increased BAG3 and HspB8 levels (in between 2 to 3 25.30
Stphanie Simon fold) in control
70.79 (19) HeLa
cells (Figure 6B), whereas expressions of their wild type counterparts had no effect.
Contrastingly,
Andre-Patrick Arrigo BAG3 and HspB8 levels were steady in p65-depleted cells
Ecole Pratique des expressing
Hautes Etudes HspB5
and SOD1 mutated forms (Figure 6B). Our results thusFlorence
indicate that protein aggregation
1 0 Manero 21.59 66.41 (13)
Readstresses
Citations
activate NFB, which in turn enhances BAG3 and HspB8 expression, two proteins
known to be involved in the stimulation of selective autophagy.
9 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 9
4 12

HspB8 and BAG3 co-localize with the protein aggregates generated by amino acid
Emeritus professor
analog,
Emeritus professorMG132 treatments or by expression of HspB5 andAdd
SOD1a mutants
new Article
Since BAG3 and HspB8 expression is upregulated via NFB upon proteotoxicity, we next
asked whether they could localize to protein aggregates. BAG3 and HspB8 localization was
Overview
thus
Contributions analyzed
Timeline by confocal
Info

immunofluorescence
Stats Scores

in untreated cells or cells submitted to

azetidine, canavanine and MG132 treatments (Figure 7 and 8, A to D). We observed a

diffuse cytoplasmic pattern of expression of BAG3 and multi-ubiquitinated
Centre Lon Brard proteins in non-
treated cells, with on
no co-localization (Figure
7A). In contrast, as was previously observed
Lyon, France
right now?(Figure 1), amino acid analogs and MG132 treatments induced the formation of cytosolic
granules
Add your current project containing
to share updates on your multi-ubiquitinated
work proteins. Of interest, we observed that those
aggregates co-localized with BAG3 as shown by nearly total overlay of BAG3 and Multi-
ubiquitin fluorescence peaks corresponding to aggregates (Figure
ones (Hsp27, 7, B to
alphaB-crystallin) andD). Additionally,
participated in the discovery
Add
2 of a crucial proteolytic machine that we called the proteasome.
square Pearsons correlation coefficients (r ) in aggregate-containing portions of the cells
were about 0.8 to 0.9 (tables, figures 7, B to D). Contrastingly, square Pearsons coefficients
were of 0.09 or 0.11 in non-treated cells or in portions of cells
Connect withdevoid of aggregates. At last,
more colleagues
we could determine that 88 to 96% of the multi-ubiquitinated aggregates
Add your other detected
social networkafter amino
acidwhat
the best way to show your peers analog or working
you've been MG132 treatments
on, create exposure forco-localized
your current with BAG3 (Figure 7 B to D). As for HspB8
updates.
(Figure 8), its distribution was rather diffuse in the cytoplasm and less abundant in the
Connect now
nucleus; moreover, it did not co-localize with multi-ubiquitin in non-treated cells (Figure 8A, r2
= 0.12). However, upon amino acid analog and MG132 treatments, HspB8 redistributed to
191 6.16k 12,280 661.97
multi-ubiquitinated granules 123
as evidenced Skills and
by channels expertise (32)
overlays, fluorescence intensity
graphs and square Pearson correlation coefficients 10
of Cell
0.7Biology 8 Heat
to 0.84. InShock Proteins HspB8
addition, 6 Apoptosis
co-
localized with more than 90% of the multi-ubiquitinated
6 aggregates
Cancer Biology 6detected (Figure
Reactive Oxygen Species 8, B to
Featured publications D). pcDNA3-HspB5WT transfected cells (HspB5wt) showed a homogeneous and diffuse
cytoplasmic
Article: localization
NF B is a central regulator ofof HspB5
protein (Figures 7E and 8E); moreover
quality BAG3 (Figure 7E) and
control in response to protein aggregation stresses via Top co-authors
HspB8modulation
autophagy (Figure 8E) distribution in those cells was similar to the one observed in non-treatedView all
Carole Kretz 32.81 220.82 (43)
cells (NT, Figures 7A and 8A), with no obvious co-localization
Claudeof BAG3
Bernard or HspB8
University Lyon 1 with HspB5
Patrick2 Arrigo Claudio Hetz Julie D Atkin Carole Kretz-Remy
(r below 0.14, see respective tables). Contrastingly, Chantal
the expression
Diaz-Latoud of HspB5R120G
28.03 126.96 (32)
Source
63 generated
0 r2 =Bernard
aggregates co-localizing with BAG3 (Figure 7F,Claude 0.71)University
and HspB8Lyon 1 (Figure 8F,
Reads2
r = Citations
0.79) proteins. Besides, this co-localization was observed in around 88 % of the
aggregates detected (Figures 7Fand 8F, tables). Similar observations were performed after
Catherine
expression of SOD1 protein in HeLa cells. BAG3 (Figure 7G) Pauland
30.45 151.49(Figure
HspB8 (16) 8G)
localization was not modified by the expression of wild type SOD1. In contrast, the proteins
1 0 Florence Manero 21.59 66.41 (13)
Readpredominantly relocalized to the inclusions generated by the of expression of SOD1G85R-
University Angers
Citations
EGFP (Figures 7H and 8H) and SOD1G93A-EGFP (Figures 7I and 8I). Indeed, about 80% to
93% of the SOD1G85R and SOD1G93A aggregates detected co-localized with BAG3 or
Chapter: Immense Cellular 2Implications Associated to Did this researcher author this publication with you?
HspB8
Small proteins
Stress Proteins (r between
Expression: Impacts0.71 to 0.86,
on Human in comparison to r2 lower than 0.14 in wtSOD1
10 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 10
4 12

expressing cells, see respective tables). Hence our results indicate that BAG3 and HspB8
Emeritus professor
redistribute
toward a vast majority of the protein aggregates generated Article
by diverse proteotoxic
stresses.
Taken together our results demonstrate that NFB is a central sensor of protein aggregation
Overview
that can modulate
protein
Info quality
Stats control

by increasing the expression of BAG3 and HspB8,
Scores
which redistribute to protein aggregates and favor their selective autophagic clearance.
Centre Lon Brard
What are you working on
DISCUSSION Apoptosis, Cancer and Development Laboratory
Lyon, France
right now?Unchecked protein misfolding and/or aggregation are the root cause of many diverse
diseases
Add your current project to share called protein
updates on conformational
your work diseases (Herczenik and Gebbink, 2008). Protein
aggregation can result in various structural appearances ranging from unordered amorphous
aggregates to highly ordered fibrils that are called amyloids.
ones (Hsp27, Here,andwe
alphaB-crystallin) studied
participated the
in the discovery
Add
involvement of NFB in the cell response to protein aggregation stress, an event also
described as proteotoxicity (Hightower, 1991). To this end, we performed treatments with
structural analogs of amino acids that generate randomly abnormal
Connect with proteins, which are
more colleagues
described to be rapidly ubiquitinated, overwhelming the chaperones
Add your otherand the proteasome
social network services to Mnd more
system
the best way to show your peers and been
what you've thus inducing
working the
on, create formation
exposure of cytosolic
for your current granules (Dasuri
updates. et al., 2011). We indeed
observed that amino acid analog treatments induced the formation of small multi-
Connect now
ubiquitinated cytosolic granules but also aggresome-like structures under more drastic
treatments (Figure 1). In this respect, it is interesting to note that canavanine treatments
191 6.16k
seem 12,280
to induce 661.97 123and bigger-sized
more frequent Skills and in
aggregates expertise (32) this may be related
the cells;
to a better incorporation into proteins of this analog of arginine, since cells were cultured in
medium devoid of arginine but not of proline. Cell treatments by MG132
6 Cancer Biology provoked
6 Reactive increased
Oxygen Species
Featured publications level of multi-ubiquitinated proteins and formation of ubiquitin-containing aggresomes as was
Article: NF B is a central
previously regulator
described of protein quality
(Hightower, 1980; Gies et al., 2010). We also induced other kinds of
proteotoxicity
autophagy modulation by expressing mutated proteins that are prone to form aggregates. We
Carole Kretz 32.81 220.82 (43)
observed the formation of perinuclear
aggregates after expression of theLyon
Claude Bernard University R120G
1 mutated
form of HspB5, which is in line with the description ofChantal
HspB5R120G
Diaz-Latoud aggregation with
28.03 126.96 (32)
Source
63 intermediate
0 filaments leading to aggresome formation Claude
containing the amyloid
Bernard University Lyon 1 oligomer
Reads Citations
(Meehan et al., 2007; Simon et al., 2007b). At last, weStphanie
expressed
Simon the
25.30G93A
70.79 and
(19) G85R
mutated form of SOD1 in HeLa cells and observed cytoplasmic and perinuclear inclusions.
Those aggregates are described to contain various proteins
or molecular complexes such as
the copper chaperone for superoxide dismutase (CCS), Hsp70 and proteasomal proteins.
1 0 Florence Manero 21.59 66.41 (13)
ReadMoreover,
Citations recent studies indicated that SOD1 aggregatesUniversity
might ofbeAngers
heterogeneous, ranging
from amorphous aggregates to amyloid fibrils (Matsumoto et al., 2005; Oztug Durer et al.,
2009). Of interest, the aggregates formed by the various stresses performed are of different
sizes.
Small StressThis is aExpression:
Proteins quite common
Impacts observation
on Human and this variability in size but also in structure was
11 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 11
4 12

illustrated by the following observation: depending on the stress induced (temperature, pH,
Emeritus professor
pressure)
Emeritus professor Add aprotein
but also of the intracellular environment (for example new concentration),
Article a
given protein can aggregate into diverse morphological/structural and toxic states (Wang et
al., 2010). Moreover, many studies describe that if the clearance systems such as UPS or
Overview autophagy
Contributions are overwhelmed,
Timeline Info the
Stats aggregated

Scores proteins accumulate and are usually gathered

at specific sites, generating larger size aggregates such as aggresomes at MTOC, to
minimize their toxic effects (Garcia-Mata et al., 1999). In thisCentre
study,
Lon we therefore had to our
Brard
disposal on
various treatments Apoptosis, Cancer and Development Laboratory
that allow the induction of diverse types of protein aggregation
Lyon, France
right now?stresses: random or selective aggregation of proteins, amorphous aggregates or amyloid
fibrils
Add your current project and updates
to share were able
on yourtowork
demonstrate that even though those aggregates are diverse in their
origin, structure and size, they all triggers NFB activation.
Of interest, MG132 is usually described as an inhibitorones
of(Hsp27,
the canonical activation
alphaB-crystallin) pathway
and participated of
in the discovery
Add
NFB via its inhibitory effect on IB degradation by the proteasome (Zanotto-Filho et al.,
2012). Surprisingly, we observed that MG132 treatment could activate NFB, suggesting the
involvement of an alternative activation pathway. And wewith
Connect indeed demonstrated that this
more colleagues
stimulation does not occur through the canonical NFB transduction pathway,
Add your other
since,
social network byto Mnd
services themore
the best way to show your peers
use what you've been working
of dominant on, create
negative exposure forwe
mutants, your current
observed that NFB activation
updates. by MG132 or mutated
HspB5 and SOD1 proteins is independent of IKK activity or Connect
IB now
phosphorylation
/degradation (Figure 2). Hence, our results suggest that NFB is a central sensor of protein
191 6.16k
aggregation stresses, which is activated via a non-canonical pathway. However, we could
12,280 661.97 123 Skills and expertise (32)
Publications observe
Reads that
Citations theImpact
intensity
Points of Full-texts
NFB activation
View statsvaries depending on the type of treatment, with
a maximal induction after heat stress and a minimal induction after overexpression of
mutated proteins prone to form aggregates. One could hypothesize that heat stress or
MG132 treatment affects all (heat stress) or a wide range (MG132) of proteins and induces
numerous
control in responseaggregates, whereas
to protein aggregation mutated
stresses via proteins induce specific and less numerousView all
Top co-authors
aggregates. Yet this explanation might not reflect reality since
Caroletoxicity of aggregates
Kretz 32.81 220.82 (43) and thus
Stphanie Simon Baptisteto
cell response Guey Malenn
those Fournier Andr-
aggregates does not only depend on their quantity but correlate with
Chantal Diaz-Latoud
their capacity to promote aberrant protein interactions. 28.03
Additionally,
126.96 (32) different types of
Source
aggregates can interact and/or sequester various types of proteins; these interactions being
Reads Citations
determined by specific sequence features and/or enrichment of disordered regions of the co-
aggregating
Article: proteins
Meet Our Associate (Olzscha
Editor: et al., 2011). Nonetheless, even if current knowledge does not
Andre-Patrick Arrigo Ecole Pratique des Hautes Etudes to stimulate
allow a direct comparison between aggregate sizes or levels and of their capacity
1 NFB
0 activity, our data highlight a new aspect of NFB Florence
function;
Maneroindeed,
21.59 NFB might be
66.41 (13)
Read Citations
considered as a primary regulator of protein aggregation stress response, therefore adding
another item to the growing list of stress responses in which NFB is involved (viral infection,
12 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 12
4 12

bacterial lipids, DNA damage, oxidative stress, ER stress, chemotherapeutic agents ) (Piva
Emeritus professor
et al.
Emeritus , 2006;
professor Tam et al., 2012). Add a new Article
We next wondered what were the consequences of protein aggregate accumulation and
NFB activation in stressed cells and first focused on autophagy. Indeed, autophagy is widely
Overview
describedTimeline
Contributions to be involved

Info inStats
the clearance

Scores of protein aggregates (Martinez-Vicente and

Cuervo, 2007; Nivon et al., 2009; Nivon et al., 2012). In this study, we demonstrated that
amino acid analog treatments increased the autophagic flux (Figures 3 and S1). Similarly, we
Centre Lon Brard
observed that the on
autophagic flux is activated upon proteasome inhibitor MG132 treatment,
Lyon, France
right now?as already observed in cultured astrocytes (Janen et al., 2010). At last, we observed that
overexpression
Add your current project to share updatesof
on SOD1G85R,
your work SOD1G93A or HspB5R120G stimulated the autophagic flux
whereas the wild type forms of those proteins did not. These results are consistent with a
previous analysis performed with SOD1G93A mutant in neuron-like
ones (Hsp27, mouse
alphaB-crystallin) cells inNSC34
and participated the discovery
Add
(Wei, 2014). As for HspB5R120G, our results are in line with the description of autophagy
activation after HspB5R120G expression in cardiomyocytes (Tannous et al., 2008). However,
they contrast with another study reporting that autophagy
Connect is inhibited
with more colleagues (altered
autophagosome maturation) in cardiomyocytes and lens from HspB5R120G knock-in
Add your other social network mouse
et al., 2013).updates.
models
the best way to show your peers (Maloyan
what you've and
been working Robbins,
on, create 2010;
exposure for Wignes
your current This discrepancy might be
explained by the duration of expression of this mutant. Indeed, in the former study a long-
Connect now
term expression of HspB5R120G was performed, whereas in the present study we performed
autophagy measurements 24 hours after the beginning of the transient expression of
191 6.16k 12,280
HspB5R120G. 661.97
Additionally, 123
HspB5
is known Skills and
to chaperone expertisefilaments
vimentin (32) that have been
10 Cell Biologywith
recently shown to suppress autophagy dynamics by interacting 8 Beclin1,
Heat Shock Proteins 6 Apoptosis
a protein crucial
for autophagy initiation (Mostowy, 2014). Thus, it could
6 be hypothesized
Cancer Biology that
6 Reactive theSpecies
Oxygen expression
Featured publications of HspB5R120G, whose chaperone activity is altered, could induce the formation of
aggregates
Article: with regulator
NF B is a central vimentin filaments
of protein qualityand provoke the release of Beclin1 that, in turn, could
control in response to protein aggregation stresses via Top co-authors
stimulate
autophagy autophagy. Nonetheless, in a long-term
modulation context, the accumulation of HspB5R120GView all
Carole Kretz 32.81 220.82 (43)
aggregates could overwhelm the autophagic process or the
Claudelysosome machinery
Bernard University Lyon 1 and alter
the autophagy-lysosome fusion process. Next, we went further and determined
Chantal Diaz-Latoud that NFB-
28.03 126.96 (32)
Source
63 deficient
0 cells submitted to those various protein aggregation stresses
Claude Bernard Universityexhibited
Lyon 1 a strong
Reads Citations
impairment of autophagy stimulation (Figure 4). Thus, evenStphanie
though autophagy
Simon was (19)
25.30 70.79 described
to be involved in the clearance of protein aggregates generated by MG132 treatments, by
expression of HspB8R120G in cardiomyocytes or by
SOD1G85R and SOD1G93A
expression in mouse neuroblastoma cell line (Kabuta et al., 2006; Janen et al., 2010;
1 0 Florence Manero 21.59 66.41 (13)
ReadPattison et
Citations al., 2011), to our knowledge, it is the first time that
connections can be performed
between activation of NFB transcription factor by those protein aggregation stresses and
stimulation of autophagy. We thus wondered if the clearance of protein aggregates was
Small Stress Proteins
modulated Expression: Impacts onautophagy
by NFB-dependent Human and analyzed protein aggregation status in NFB-
13 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 13
4 12

deficient cells submitted to amino acid analogs, MG132 and expression of HspB5 and SOD1
Emeritus professor
mutants.
We observed a higher accumulation of protein aggregates Article
in those cells than in
control cells (Figure 5), leading to the conclusion that NFB-dependent autophagy is involved
in the clearance of various kinds of protein aggregates generated by diverse aggregation
Overview
stresses. Timeline
Contributions
Info
Stats
Scores
To determine how NFB could modulate autophagy in response to proteotoxicity, we focused

Centre Lon Brard
on BAG3/HspB8 complex. Indeed, BAG3 co-chaperone is described as a molecular switch
Lyon, France
that addresses proteins to proteasomal or autophagic degradation; it is also found to increase
right now?
autophagic activity (Carra, 2009; Gamerdinger et al., 2011; Rapino et al., 2014). Moreover,
BAG3 binds to chaperones
such as Hsp70 and HspB8, which allow recognition and binding
About
to misfolded proteins (Carra et al., 2008a; Chen et Ial.,
have 2013). BAG3
studied Heat is also
shock proteins reported
(Hsps), particularly to i)
the small
facilitate the retrograde transport of misfolded
Add proteins to the MTOC, ii) to form aggresome
(Gamerdinger et al., 2009) and iii) to promote both autophagy induction via its binding to Atg3
(Chen et al., 2013) and selective autophagy through its interaction with p62 (Gamerdinger et
al., 2009). Accordingly, the BAG3/HspB8 complexConnect
was with more colleagues
reported to selectively activate
such as Htt43Q Add
or your (Carra et al.,
other social network services to Mnd more
autophagic
Use your proMle overview page removal
to present yourself of aggregated
and your research. proteins
Customizing your proMle is SOD1G93A
2008a; Crippa et al., 2013); Interestingly, in human, BAG3
updates.
mutations induce severe
myofibrillar myopathy associated to cardiomyopathy and neuropathy,
Connectwhereas
now HspB8
mutations are associated to neuropathy (Fontaine et al., 2006; Selcen et al., 2009;
191 Konersman et al., 2015). At last, we previously demonstrated that NFB activates autophagy
6.16k 12,280 661.97 123 Skills and expertise (32)
Publications in
Reads response
Citations to heat
Impactshock
Points treatments
Full-texts via the
View upregulation of BAG3 and HspB8 expression. In
stats
this study, we observed that amino acid analogs or MG132 treatments but also expression of
HspB5R120G, SOD1G85R and SOD1G93A induced an increase of BAG3 and HspB8 levels
(Figure 6). Hence, our results corroborate the increased transcription of bag3 and hspb8
observed
control after
in response MG132
to protein treatments
aggregation (Du
stresses via et al., 2009; Minoia et al., 2014) and the increased
Top co-authors View all
levels of BAG3 and HspB8 detected in muscles of SOD1G93A
Carole Kretz knock-in mouse
32.81 220.82 (43) models
Stphanie
(CrippaSimonet al., Guey
Baptiste Malenn
2013). Fournier Andr-
Importantly, to our knowledge, our study reports for the first time a
Chantal Diaz-Latoud
modulation of BAG3 and HspB8 levels after protein aggregation
28.03 stresses
126.96 (32)such as amino acid
Source
analog treatments or expression of the R120G mutated form of HspB5. Of interest, in this
Reads Citations
latter context it has been reported that BAG3 interacts withStphanie Simon 25.30 70.79 (19)
HspB5R120G and suppresses its
aggregation
Article: and Editor:
Meet Our Associate toxicity (Hishiya et al., 2011). Similarly, transfection of HspB8 in
cardiomyocytes is described to block the amyloid oligomerEcole Pratique by
formed des Hautes Etudes
HspB5R120G (Sanbe
1 et al.,
0 2007). We next investigated if increased BAG3Florence
andManero
HspB8 levels
21.59 66.41induced
(13) by
Read Citations
proteotoxicity were dependent on NFB activity, similarly to what we showed after a heat
shock treatment (Nivon et al., 2012). Impaired stimulation of BAG3 and HspB8 expression
was Immense
Chapter: observed afterImplications
Cellular protein aggregation
Associated to Did this researcher author
stresses in p65-depleted cells,thisindicating
publication with
thatyou?NFB
14 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 14
4 12

is an important regulator of BAG3 and HspB8 expression in response to amino acid analog
Emeritus professor
andprofessor
Emeritus MG132 Add a new
treatments or to expression of HspB5R120G, SOD1G85R andArticle
SOD1G93A. As
for MG132 treatment, these results are consistent with a previous study reporting that the
heat shock response, the unfolded protein response and the NFB signaling pathway all
Overview
participate
Contributions in BAG3 upregulation
Timeline Info Stats uponScores

proteasome

inhibition (Minoia et al., 2014); indeed,we
observed that the inhibition of BAG3 upregulation in NFB-deficient cells is not complete,
suggesting the involvement of other regulators. Taken together,
Centreour
Lon data
Brard indicate that NFB
deficiency impairson
BAG3/HspB8 upregulation after variousLyon,
protein
France aggregation stresses,
right now?which is associated with a defective autophagic process and an accumulation of protein
Add your current project to share updates
aggregates. on your work this
Interestingly, process seems to be rather restricted to BAG3 and HspB8
partners. Indeed, we could not detect any modulation of the transcript levels of other known
players of protein aggregate clearance
Add
such as Alfy,
onesNBR1 or optineurin
(Hsp27, alphaB-crystallin) and in a microarray
participated in the discovery
analysis performed in control and p65-depleted HeLa cells submitted to heat stress, which is
an inducer of the non-canonical NFB pathway (unpublished results).
At last, we checked whether the upregulated BAG3 and HspB8
Connect could
with more associate with protein
colleagues
Add HspB8
aggregates and observed a strong co-localization of BAG3 and your other social
withnetwork services to Mnd more
the aggregates
generated
the best way to show your peers what you'veby amino
been acid
working on, analog
create exposureand MG132
for your current treatments or byupdates.
the expression of HspB5 and
SOD1 mutated forms. These observation are in line with the co-localization of BAG3 with
Connect now
multi-ubiquitinated aggregates described upon MG132 treatments (Minoia et al., 2014), the
co-localization of overexpressed BAG3 and HspB5R120G in HEK293 cells (Hishiya et al.,
191 6.16k 12,280 661.97 123
Publications Reads
2011) Citations
or the association
Impact Points
of Full-texts
BAG3 andView
SOD1G85R
stats
aggregates (Gamerdinger et al., 2011)
and the co-localization of HspB8 and SOD1G93A in mice models (Crippa et al., 2010),
indicating that the NFB-dependent upregulation of6BAG3

Cancer Biology 6 Reactive Oxygen Species
and HspB8 expression might
facilitate the recruitment of BAG3/HspB8 to protein aggregates and strengthen the role of
BAG3 and HspB8 in the selective removal of aggregates via chaperone associated selective
autophagy
autophagymodulation
(CASA). Carole Kretz 32.81 220.82 (43)
Chantal
All together, our results further highlight the critical and Diaz-Latoud
central
role played by NFB
28.03 126.96 (32)
Source
63 transcription
0 Claude Bernard University Lyon 1
factor in protein quality control. Our data indicate that NFB is a general sensor
Reads Citations
Stphanie Simon
of protein aggregation stress that can be activated by random 25.30 70.79
or selective (19)
aggregation of
proteins,
Article: Meet Ouramorphous aggregates
Associate Editor: or amyloid fibrils. Its activation induces an on-demand
compensatory mechanism that is the upregulation of the BAG3-HspB8
Ecole Pratique des complex,
Hautes Etudes which in turn
1 associates
0 with protein aggregates and stimulates selective
Florenceautophagy.
Manero 21.59 This
66.41 mechanism
(13)
Read Citations
allows the clearance of toxic protein aggregates and thus helps the cell to recover from
proteotoxicity. Hence this molecular pathway may impact therapies for protein conformational
diseases,
Chapter: Immensenotably cardiac dysfunction,
Cellular Implications Associated to cataract formation or neurodegenerative
Did this researcher author this publication withdiseases
you?
15 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 15
4 12

associated to proteasome impairment, myofibrillar myopathies associated with HspB5
Emeritus professor
mutations
Emeritus professor Addmutations.
or familial amyotrophic lateral sclerosis linked to SOD1 a new Article
MATERIALS AND METHODS

Overview Contributions Timeline Info Stats Scores
Cell culture and cell lines

HeLa cells were grown at 37C and 5% CO2 in DMEM supplemented with 10%
Centre Lon Brard
decomplemented FCS, 1 g/ml fungizone and 50 U/ml penicillin/streptomycin (Nivon
Apoptosis, Cancer and Development et al.,
Laboratory
2009). For amino acid analogs treatments, cells were first Lyon, France
right now? cultivated during 1 h in DMEM
without L-Arginine (PAN Biotec) supplemented with 10% dialyzed fetal calf serum. Then,
cells
and get noticed for your were incubated with 5
research. to 15 mM of L-canavanineAbout
sulfate (abbreviated canavanine) or L-
azetidine-2-carboxylic acid (abbreviated azetidine). For E64d
I have studiedand pepstatin
Heat shock A studies,
proteins (Hsps), 5 the
particularly M small
Add
of theses inhibitors were added concomitantly with the treatments
of a crucial proteolyticor just that
machine after transfections
we called the proteasome.
and were not removed until cellular protein extraction. HeLa cells expressing shRNA directed
against the p65 subunit of NF-B (p65-KD#2) have been already described (Nivon et al.,
2009). It was shown in previous studies that p65-KD#2 cell line behaves like other p65-
Use your proMle overview page to presentclones
deficient yourself and your research.
obtained inCustomizing
the team, yourin
proMle is
terms of growth, autophagic activity orachievements
responseandto
colleagues and share your
projects, and start building heat stress;
your network. similarly, control HeLa cells used in this study have the same behavior than
HeLa cells expressing single-mutated or scrambled-mutated shRNAConnect now
directed against p65
(Nivon et al., 2009; Nivon et al., 2012). Stable HeLa transfectants expressing GFP-LC3 have
191 been already
6.16k 12,280 described 123 et al., 2005).
661.97 (Bampton Skills and expertise (32)
Publications Reads
Reagents and plasmids
Citations Impact Points Full-texts View stats
E64d, pepstatin A, L-azetidine-2-carboxylic acid, L-canavanine sulfate, Hoechst 33258 and

Triton X-100 were from Sigma. MG132 was from Merck. Mouse monoclonal antibodies used
were directed against: Actin, clone C4 (Millipore); p62/SQSTM1, clone 3/p62 lck ligand
control in response
(Becton to proteinBiosciences);
Dickinson aggregation stresses via
multi-ubiquitin, clone
TopFK1 (MBL); EGFP, clone 1GFP-2A5View all
co-authors
(Euromedex); HspB8, Carole Kretz 32.81 220.82
clone (43)1B6.1-3G4
Mathieu Nivon Loc Fort Pascale Mullerclone 3C5 (Abnova); alphaB crystalline/HspB5,
Emma Richet Claude Bernard University Lyon 1
(Enzo).
Patrick Anti-LC3
Arrigo Claudio (Sigma,
Hetz Julie D Atkin #L7543), BAG3 (Abcam, #Ab47124), HspB8 (Abnova, #PAB15424)
Carole Kretz-Remy
Chantal Diaz-Latoud
and anti-pericentrin (Abcam, #Ab4448) were rabbit polyclonal 28.03 antibodies.
126.96 (32) pcDNA3-BCWT
Source
and pcDNA3-BCR120G
Reads Citations are expression vectors for wild type HspB5 protein or its R120G
mutated form (Simon et al., 2007b); pCDNA3.1(+) (Invitrogen) wasCrteil
Universit Paris-Est used asMarne
Val de the... empty
Article: Meet Our control
plasmid Associate Editor:
for those constructs. pEGFP-SOD1 WT G85R
Catherine,Paul pEGFP-SOD1
30.45 151.49 (16) and
G93A
pEGFPSOD1 are expression vectors for wild type hSOD1-EGFP and its mutated forms
1 hSOD1
0 G85R-EGFP and hSOD1G93A -EGFP; pEGFP-N1 (Clontech)
Florence Manero 21.59 66.41 (13)
was used as the empty
Read Citations
plasmid control for those constructs (Turner et al., 2005). pLXSN-IBM plasmid allows the
expression of a dominant negative mutant of IB in which Ser32, Ser36, Ser283, Ser288
and Ser293 phosphorylation sites are mutated
to alanine (Kretz-Remy et al., 2001). pRK5-
16 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 16
4 12

IKK(K44A)-C-Flag plasmid encodes a dominant negative mutant of IKK (Kretz-Remy et al.,
Emeritus professor
2001).
Emeritus pNFBluc
professor was from Clontech. Add a new Article
Transfection
Cells were seeded the day before transfection at a density of 5.105 cells/60 mm dishes. They
Overview
were
Contributions thenTimeline
transfectedInfo

with 5 Stats
g of theScores

desired plasmid according to the ExGen 500 reagent

procedure.
Luciferase assay Centre Lon Brard
What are you
Stableworking
transfectanton
HeLa cells expressing pNFBluc (Clontech) were submitted to various
Lyon, France
right now?treatments or transfections. Thereafter, they were resuspended into PBS and luciferase
activity
Add your current project was
to share quantified
updates using
on your work the steady-glo luciferase assay system according to the
manufacturers instructions (Promega). The intensity About
of light produced was quantified with a
Victor3 Luminometer (Perkin Elmer). The related lightones
units produced
(Hsp27, wereand
alphaB-crystallin) reported
participatedto 50discovery
in the g
Add
of total cellular proteins
Gel electrophoresis and immunoblotting
Briefly, 10 g of total protein extracts were loaded onto SDS-PAGE.
Connect After electrophoresis,
proteins were transferred onto Protran BA85 nitrocellulose membrane (Whatmann).
Add your other social network services Blots
to Mnd more
werewhat
the best way to show your peers hybridized withon,primary
you've been working antibodies
create exposure and
for your current horseradish-peroxidase-conjugated
updates. secondary
antibodies (Bio Rad, #170-65-16 and #170-6515) and revealed with ECL blotting detection
Connect now
reagents (Amersham) or Clarity western ECL substrate (Bio Rad). Western blot imaging was
performed with Chemidoc MP (Bio Rad) based on CCD detection technology. Image capture
191 6.16k 12,280 of661.97
and analysis 123were controlled
western blots Skills and
by Image expertise
Lab (32)
software. Quantification of
western blots was performed by Image J software.
Filter trap assay 6 Cancer Biology 6 Reactive Oxygen Species
Featured publications SDS-insoluble aggregates were analyzed by filter trap analysis as previously described
Article:
(NivonNF B et
is aal.,
central regulator
2012). of protein
Briefly, cellsquality
were scraped in 2% SDS-FTA buffer (FTA: 150 mM NaCl,
50 mM
autophagy DTT, 10 mM Tris-HCl, pH 8). Next, samples were homogenized by three passages
modulation
Carole Kretz 32.81 220.82 (43)
through 25G needle. 2.5 g of protein
extracts were diluted byBernard
Claude a factor of 2Lyon
University to1 8 (1, 1:2, 1:4
and 1:8) and applied with mild suction into a slot blot apparatus onto a 0.22
Chantal Diaz-Latoud m protran BA83
28.03 126.96 (32)
Source
63 nitrocellulose
0 membrane (Schleicher and Schuell) pre-washed withUniversity
Claude Bernard 0.1% LyonSDS-FTA
1 buffer.
Reads Citations
Then the membrane was washed with 0.1%SDS-FTA buffer
Stphanieand
Simon0.1%Tween-TBS
25.30 70.79 (19) buffer
(TBS: 20 mM Tris-HCl, pH7.6, 137 mM NaCl) and processed for immunoblotting. To note,
equivalent expression levels of wt and mutant constructs for
HspB5 and SOD1 was checked
on western blots of total cell lysates, in parallel experiments.
1 0 Florence Manero 21.59 66.41 (13)
ReadFluorescence microscopy analysis University of Angers
Citations
Non treated, treated or transfected cells were grown on glass coverslips followed by various
recovery periods, rinsed in cold PBS and either fixed for 30 min in 2% paraformaldehyde pH
7.4Stress
Small (EGFP expressing
Proteins Expression: cells)
Impactsor
on fixed
Humanfor 10 min with cold methanol and permeabilized with
17 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 17
4 12

0.1% triton X-100. Cells were stained with primary antibodies (MultiUb, HspB5, HspB8,
Emeritus professor
BAG3,
Emeritus pericentrin,
professor LC3) and fluorescent secondary antibodiesAdd a new
(goat Article
anti-mouse Alexa Fluor
568 or 488 nm and goat anti-rabbit Alexa Fluor 488 or 568 nm (Life technologies)). Hoechst
33258 reagent was used to stain nuclei (5 min, 1 ng/ml). For quantification of aggregation,
Overview cells
Contributions were photographed
Timeline Info with
Stats a Zeiss

Scores axiovert 200M or a Zeiss Axio Imager Z1

photomicroscope (x40 objective). Images were digitized with a camera (Coolsnap HQ2;
Roper scientific) and acquired with Metavue Imaging system.Centre
TheLon
number
Brard
of cells containing
aggregates onor larger than 3 m was quantified 3Apoptosis,
smaller timesCancer
on and
100Development Laboratory
cells. For co-
Lyon, France
right now?localization experiments, z-stacks were performed with a confocal Zeiss LSM 510 META (x63
objective).
Add your current project Co-localization
to share updates on your work was quantified by measuring, on z-stacks, Pearsons correlation
and get noticed for your research. Aboutwith Fijis JACoP plugin, on around
coefficients and geometrical center based co-localization
20 cells for each experimental conditions. ones (Hsp27, alphaB-crystallin) and participated in the discovery
Add
Statistics of a crucial proteolytic machine that we called the proteasome.
The unpaired Students t test was applied to compare results between two sample groups.
Differences between groups were considered statistically significant when P-values were less
than 0.05. *P<0.05, **P<0.01, ***P<0.001. Add your other social network services to Mnd more
ACKNOWLEDGEMENTS Connect now
We wish to thank Dominique Guillet for excellent technical assistance, Ludivine Walter for
critical reading of the manuscript, Charlotte Scholts for her help with confocal microscopy
191 6.16k 12,280 661.97 123
Publications and
Reads Berangre
Citations Pinan-Lucarre
Impact Points and Julien
Full-texts Falk
View stats for their help with co-localization analysis.

CONTRIBUTIONS FROM OTHERS
Mathieu Nivon, Loc Fort, Pascale Muller and Carole Kretz-Remy performed most of the
experiments.
control Emmaaggregation
in response to protein Richet, stresses
Baptistevia Guey, Malenn Fournier and Stphanie SimonView all
Top co-authors
contributed to preliminary results. Carole Kretz-Remy supervised
the studies,
Carole Kretz 32.81 220.82 (43) helped to
design and interpret experiments and
wrote the paper. All the authors discussed the results
Chantal Diaz-Latoud
and performed critical reading of the manuscript. 28.03 126.96 (32)
Source
Reads Citations
FUNDING SOURCES
ThisMeet
Article: work
Our was supported
Associate Editor: by the Ligue contre le Cancer, comit du Rhne; and the Ligue
contre le Cancer, comit de Savoie; and the Bonus Qualit Recherche from Universit
1 Claude
0 Bernard Lyon I; and the Centre National de la Recherche
Florence Manero 21.59 66.41
Scientifique (13) and the
to C.K;
Read Citations
Millennium Institute No. P09-015-F, FONDAP program 15150012 to C.H . M.N. was
supported by a doctoral fellowship from the French Department of research; and the
Did this researcher author this publication with you?
18 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 18
4 12

Fondation pour la Recherche Mdicale. S.S was supported by a post-doctoral fellowship from
Emeritus professor
theprofessor
Emeritus Association Franaise contre les Myopathies/Tlthon. Add a new Article
REFERENCES
Overview
Info
Stats
Scores
Bampton, E.T., Goemans, C.G., Niranjan, D., Mizushima, N., and Tolkovsky, A.M. (2005).
The dynamics of autophagy visualized in live cells: from autophagosome formation to
fusion with endo/lysosomes. Autophagy 1, 23-36. Centre Lon Brard
What are you2012, working
673290.
on
Behrends, C., and Fulda, S. (2012). Receptor proteins in selective Apoptosis,autophagy. Int J Cell
Cancer and Development Biol
Laboratory
Lyon, France
right now?Carra, S. (2009). The stress-inducible HspB8-Bag3 complex induces the eIF2alpha kinase
pathway: implications for protein quality control and viral factory degradation? Autophagy
Add your current project5, to share updates on your work
428-429.
Carra, S., Seguin, S.J., Lambert, H., and Landry, About J. (2008a). HspB8 chaperone activity
toward poly(Q)-containing proteins depends on itsI have association withproteins
studied Heat shock Bag3, a stimulator
(Hsps), particularly theof
small
macroautophagy. J Biol Chem 283,Add 1437-1444. ones (Hsp27, alphaB-crystallin) and participated in the discovery
Carra, S., Seguin, S.J., and Landry, J. (2008b). HspB8 of aand
crucialBag3:
proteolytic
a machine that we called the
new chaperone proteasome.
complex
targeting misfolded proteins to macroautophagy. Autophagy 4, 237-239.
Chen, B., Retzlaff, M., Roos, T., and Frydman, J. (2011). Cellular strategies of protein quality
control. Cold Spring Harb Perspect Biol 3, a004374.
Show your career's best Chen, Y., Yang, L.N., Cheng, L., Tu, S., Guo, S.J., Le, H.Y., Xiong, Q., Mo, R., Li, C.Y.,
Jeong, J.S., Jiang, L., Blackshaw, S., Bi, L.J., Zhu, H., Tao, Add
your and
S.C., other social
Ge,network services Bcl2-
F. (2013). to Mnd more
associated athanogene 3 interactome analysis reveals colleagues

a new and share your achievements and
role in modulating
proteasome activity. Mol Cell Proteomics 12, 2804-2819.
updates.
Crippa, V., Boncoraglio, A., Galbiati, M., Aggarwal, T., Rusmini, P., Giorgetti, Connect now E., Cristofani,
R., Carra, S., Pennuto, M., and Poletti, A. (2013). Differential autophagy power in the
spinal cord and muscle of transgenic ALS mice. Front Cell Neurosci 7, 234.
Crippa, V., Sau, D., Rusmini, P., Boncoraglio, A., Onesto, E., Bolzoni, E., Galbiati, M.,
191 6.16k Fontana,12,280 E., 661.97 Marino, M., Carra, 123 S., Bendotti, C., De Biasi,
Skills S., and(32)Poletti, A. (2010). The
and expertise
Publications Reads small Citations
heat shock Impact Points
protein B8 Full-texts
(HspB8) View stats
promotes autophagic removal of misfolded proteins
involved in amyotrophic lateral sclerosis (ALS). Hum Mol Genet 19, 3440-3456.
Dahlmann, B. (2007). Role of proteasomes in disease.6BMC Biochem

Cancer Biology 8 Suppl 1, S3.
6 Reactive Oxygen Species
Dasuri, K., Ebenezer, P.J., Uranga, R.M., Gavilan, E., Zhang, L., Fernandez-Kim, S.O.,
Bruce-Keller, A.J., and Keller, J.N. (2011). Amino acid analog toxicity in primary rat
Article:neuronal and regulator
NF B is a central astrocyte cultures:
of protein qualityimplications for protein misfolding and TDP-43 regulation.
control JinNeurosci
response toRes 89aggregation
protein , 1471-1477. stresses via Top co-authors View all
autophagy
Du, Z.X., modulation
Zhang, H.Y., Meng, X., Gao, Y.Y., Zou, R.L., Liu, B.Q., Guan, Y., and Wang, H.Q.
Carole Kretz 32.81 220.82 (43)
Mathieu (2009).
Nivon Loc Proteasome
Fort Pascale Mullerinhibitor
Emma Richet
MG132 induces BAG3 expressionClaude Bernard through activation
University Lyon 1 of heat
shock factor 1. J Cell Physiol 218, 631-637.
Edington, B.V., Whelan, S.A., and Hightower, L.E. (1989). Chantal Diaz-Latoud
Inhibition of heat shock (stress)
28.03 126.96 (32)
Source
63 protein
0 induction by deuterium oxide and glycerol: additional support for the abnormal
Reads
protein hypothesis of induction. J Cell Physiol 139, 219-228.
Citations
Elliott, J.L., Der Perng, M., Prescott, A.R., Jansen, K.A., Koenderink, Stphanie SimonG.H., 25.30and Quinlan,
70.79 (19) R.A.
(2013). The specificity of the interaction between alphaB-crystallin and desmin filaments
Article:and
Meet its
Ourimpact
Associateon filament aggregation and cell viability. Philos Trans R Soc Lond B Biol
Editor:
Andre-Patrick 368, 20120375.
Sci Arrigo Ecole Pratique des Hautes Etudes
Fontaine, J.M., Sun, X., Hoppe, A.D., Simon, S., Vicart, P., Welsh, M.J., and Benndorf, R.
1 0
(2006). Abnormal small heat shock protein interactions Florence Manero neuropathy-associated
involving 21.59 66.41 (13)
Citations (HSPB8) mutants. FASEB J 20, 2168-2170.
Read HSP22
Gamerdinger, M., Hajieva, P., Kaya, A.M., Wolfrum, U., Hartl, F.U., and Behl, C. (2009).
Protein quality control during aging involves recruitment of the macroautophagy pathway
by BAG3. The EMBO journal 28, 889-901. Did this researcher author this publication with you?
19 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 19
4 12

Gamerdinger, M., Kaya, A.M., Wolfrum, U., Clement, A.M., and Behl, C. (2011). BAG3
Emeritus professor
mediates chaperone-based aggresome-targeting and selective Add a autophagy
new Article of misfolded
proteins. EMBO Rep 12, 149-156.
Emeritus professor
Garcia-Mata, R., Bebok, Z., Sorscher, E.J., and Sztul, E.S. (1999). Characterization and
dynamics of aggresome formation by a cytosolic GFP-chimera. J Cell Biol 146, 1239-
1254.
Overview
Gies, E., Timeline
Contributions
Wilde, I., Winget,

Info
J.M.,
Stats
Brack, M., Rotblat, B., Novoa, C.A., Balgi, A.D., Sorensen,
Scores

P.H., Roberge, M., and Mayor, T. (2010). Niclosamide prevents the formation of large
ubiquitin-containing aggregates caused by proteasome inhibition. PLoS One 5, e14410.
Goldberg, A.L. (2003). Protein degradation and protection Centre against misfolded or damaged
Lon Brard
proteins. Nature 426, 895-899. Apoptosis, Cancer and Development Laboratory
Haigis, M.C., and Yankner, B.A. (2010). The aging stress response. Lyon, France Mol Cell 40, 333-344.
right now?Hayden, M.S., and Ghosh, S. (2004). Signaling to NF-kappaB. Genes Dev 18, 2195-2224.
Herczenik, E., and Gebbink, M.F. (2008). Molecular and cellular aspects of protein misfolding
Add your current projectand to share updates FASEB
disease. on your workJ 22, 2115-2133.
Hightower, L.E. (1980). Cultured animal cells exposedAbout to amino acid analogues or puromycin
rapidly synthesize several polypeptides. J Cell Physiol I have102,
studied407-427.
Heat shock proteins (Hsps), particularly the small
Hightower, L.E. (1991). Heat shock, stress Add
ones (Hsp27, alphaB-crystallin)
proteins, chaperones, and participated Cell
and proteotoxicity. 66,
in the discovery
191-197.
Hishiya, A., Salman, M.N., Carra, S., Kampinga, H.H., and Takayama, S. (2011). BAG3
directly interacts with mutated alphaB-crystallin to suppress its aggregation and toxicity.
PLoS One 6, e16828. Connect with more colleagues
Show your career's best Holmes, W.M., Klaips, C.L., and Serio, T.R. (2014). Defining the limits: Protein aggregation
Use your proMle overview page to and toxicity
present yourselfinandvivo. Crit Rev
your research. Biochem
Customizing MolisBiol 49, 294-303. Add your other social network services to Mnd more
your proMle
Janen,
the best way to show your peers S.B.,been
what you've Chaachouay,
working on, create H., andforRichter-Landsberg,
exposure your current C. (2010). Autophagy is activated by
updates.
proteasomal
projects, and start building your network. inhibition and involved in aggresome clearance in cultured astrocytes. Glia
58, 1766-1774. Connect now
Kabuta, T., Suzuki, Y., and Wada, K. (2006). Degradation of amyotrophic lateral sclerosis-
linked mutant Cu,Zn-superoxide dismutase proteins by macroautophagy and the
proteasome. J Biol Chem 281, 30524-30533.
191 6.16k
Kim, J., 12,280
Lee, H.,661.97 Lee, J.H., Kwon, 123 D.Y., Genovesio, A., Fenistein,
Skills and expertiseD.,(32)
Ogier, A., Brondani, V.,
Publications Reads andCitations Grailhe,Impact R. Points
(2014).Full-texts View stats oligomerization, and aggregation of human
Dimerization, 10 Cell Biology 8 Heat Shock Proteins 6 Apoptosis
amyotrophic lateral sclerosis copper/zinc superoxide dismutase 1 protein mutant forms in
live cells. J Biol Chem 289, 15094-15103. 6 Cancer Biology 6 Reactive Oxygen Species
Konersman, C.G., Bordini, B.J., Scharer, G., Lawlor, M.W., Zangwill, S., Southern, J.F.,
Amos, L., Geddes, G.C., Kliegman, R., and Collins, M.P. (2015). BAG3 myofibrillar
Article:myopathy
NF B is a centralpresenting
regulator of with cardiomyopathy.
protein quality Neuromuscul Disord 25, 418-422.
Kretz-Remy,
control in response toC., Bates,
protein E.E.,stresses
aggregation and Arrigo,
via A.P. (1998). Amino acid analogs activate NF-View all
Top co-authors
autophagykappaBmodulation
through redox-dependent IkappaB-alpha degradation by the proteasome without
Carole Kretz 32.81 220.82 (43)
Mathieu apparent
Nivon Loc FortIkappaB-alpha
Pascale Muller Emma Richet
phosphorylation. Consequence ClaudeonBernard
HIV-1 longLyonterminal
University 1 repeat
activation. J Biol Chem 273, 3180-3191.
Kretz-Remy, C., Munsch, B., and Arrigo, A.P. (2001). NFkappa Chantal Diaz-Latoud
B-dependent transcriptional
28.03 126.96 (32)
Source
63 activation
0 during heat shock recovery. Thermolability of the NF-kappaB.Ikappa
Claude Bernard University Lyon 1 B complex.
Reads J Biol Chem 276, 43723-43733.
Citations
Kuma, A., Hatano, M., Matsui, M., Yamamoto, A., Nakaya, H., Simon
Stphanie Yoshimori,
25.30 T.,
70.79Ohsumi,
(19) Y.,
Tokuhisa, T., and Mizushima, N. (2004). The role of autophagy during the early neonatal
Article:starvation
Meet Our Associate
period. Nature 432, 1032-1036.
Editor:
Li, N., Arrigo
Andre-Patrick and Karin, M. (1998). Ionizing radiation and short wavelength Ecole Pratique des UV activate
Hautes Etudes NF-kappaB
through two distinct mechanisms. Proc Natl Acad Sci U S A 95, 13012-13017.
1 Maloyan, 0 A., and Robbins, J. (2010). Autophagy in desmin-related Florence Manero 21.59 66.41 (13)
cardiomyopathy: thoughts
the halfway point. Autophagy 6, 665-666.
Read atCitations
Martinez-Vicente, M., and Cuervo, A.M. (2007). Autophagy and neurodegeneration: when the
cleaning crew goes on strike. Lancet Neurol 6, 352-361.
20 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 20
4 12

Matsumoto, G., Stojanovic, A., Holmberg, C.I., Kim, S., and Morimoto, R.I. (2005). Structural
Emeritus professor
properties and neuronal toxicity of amyotrophic lateral Add sclerosis-associated
a new Article Cu/Zn
superoxide dismutase 1 aggregates. J Cell Biol 171, 75-85.
Emeritus professor
Meehan, S., Knowles, T.P., Baldwin, A.J., Smith, J.F., Squires, A.M., Clements, P., Treweek,
T.M., Ecroyd, H., Tartaglia, G.G., Vendruscolo, M., Macphee, C.E., Dobson, C.M., and
Carver, J.A. (2007). Characterisation of amyloid fibril formation by small heat-shock
Overview Contributions
chaperone Timeline
proteins Info
human Stats
alphaA-, alphaB- and R120G alphaB-crystallins. J Mol Biol
Scores
372, 470-484.

Mehrpour, M., Esclatine, A., Beau, I., and Codogno, P. (2010). Autophagy in health and
disease. 1. Regulation and significance of autophagy: an overview. Am J Physiol Cell
Centre Lon Brard
Physiol 298, C776-785. Apoptosis, Cancer and Development Laboratory
Minoia, M., Boncoraglio, A., Vinet, J., Morelli, F.F., Brunsting, J.F., Poletti, A., Krom, S.,
Lyon, France
right now? Reits, E., Kampinga, H.H., and Carra, S. (2014). BAG3 induces the sequestration of
proteasomal clients into cytoplasmic puncta: implications for a proteasome-to-autophagy
Add your current projectswitch.to share updates
Autophagy on your10,work1603-1621.
Mostowy, S. (2014). Multiple roles of the cytoskeleton in bacterial autophagy. PLoS Pathog
10, e1004409. I have studied Heat shock proteins (Hsps), particularly the small
Nivon, M., Abou-Samra, M., Richet, E., ones (Hsp27, alphaB-crystallin) and participated in the discovery
AddGuyot, B., Arrigo, A.P., and Kretz-Remy, C. (2012).
NF-kappaB regulates protein quality control after ofheat a crucial proteolytic machine that we called the proteasome.
stress through modulation of the
BAG3-HspB8 complex. Journal of cell science 125, 1141-1151.
Nivon, M., Richet, E., Codogno, P., Arrigo, A.P., and Kretz-Remy, C. (2009). Autophagy
activation by NFkappaB is essential for cell survival after heat shock. Autophagy 5, 766-
Show your career's best783.
Olzscha, H., Schermann, S.M., Woerner, A.C., Pinkert, S., Add
your other
Hecht, M.H.,social network servicesG.G.,
Tartaglia, to Mnd more
the best way to show your peersVendruscolo, M., on,
what you've been working Hayer-Hartl,
create exposure for M.,
your Hartl,
current F.U., and Vabulas, updates.
R.M. (2011). Amyloid-like
projects, and start building youraggregates
network. sequester numerous metastable proteins with essential cellular functions. Cell
144, 67-78. Connect now
Oztug Durer, Z.A., Cohlberg, J.A., Dinh, P., Padua, S., Ehrenclou, K., Downes, S., Tan, J.K.,
Nakano, Y., Bowman, C.J., Hoskins, J.L., Kwon, C., Mason, A.Z., Rodriguez, J.A.,
Doucette, P.A., Shaw, B.F., and Valentine, J.S. (2009). Loss of metal ions, disulfide
191 6.16k reduction12,280 and 661.97
mutations123 related to familial ALS Skillspromote
and expertise formation
(32) of amyloid-like
Publications Reads aggregates
Citations Impactsuperoxide
from Points Full-texts
dismutase.View stats
PLoS One10 4, e5004.
Cell Biology 8 Heat Shock Proteins 6 Apoptosis
Pattison, J.S., Osinska, H., and Robbins, J. (2011). Atg7 induces basal autophagy and
rescues autophagic deficiency in CryABR120G cardiomyocytes. 6 Cancer Biology Circ 109,Species
ResOxygen
6 Reactive 151-160.
Piva, R., Belardo, G., and Santoro, M.G. (2006). NF-kappaB: a stress-regulated switch for
cell survival. Antioxid Redox Signal 8, 478-486.
Rapino,
Article: NF B is F., Jung,
a central M., ofand
regulator proteinFulda,
quality S. (2014). BAG3 induction is required to mitigate
control proteotoxicity
in response to protein viaaggregation
selective autophagy
stresses via following inhibition of constitutive proteinView all
Top co-authors
autophagy modulation pathways. Oncogene 33, 1713-1724.
degradation Carole Kretz 32.81 220.82 (43)
Mathieu Nivon LocK.J.,
Rodgers, Fort Pascale
and Muller Emma Richet
Shiozawa, N. (2008). Misincorporation Claude ofBernard
amino acidLyonanalogues
University 1 into
proteins by biosynthesis. Int J Biochem Cell Biol 40, 1452-1466.
Rosen, D.R., Siddique, T., Patterson, D., Figlewicz, D.A., Chantal Sapp, Diaz-Latoud
P., Hentati, A., Donaldson,
28.03 126.96 (32)
Source
63 D., 0 Goto, J., O'Regan, J.P., Deng, H.X., and et al. (1993). Mutations
Claude Bernard Universityin Cu/Zn
Lyon 1 superoxide
Reads dismutase
Citations gene are associated with familial amyotrophic lateral sclerosis. Nature 362, 59-
62. Stphanie Simon 25.30 70.79 (19)
Rubinsztein, D.C. (2006). The roles of intracellular Universit Paris-Est Crteil Val de Marne ...
protein-degradation pathways in
Article:neurodegeneration.
Meet Our Associate Editor: Nature 443, 780-786. Catherine Paul 30.45 151.49 (16)
Sanbe,Arrigo
Andre-Patrick A., Yamauchi, J., Miyamoto, Y., Fujiwara, Y., Murabe, M., des
Ecole Pratique and Tanoue,
Hautes Etudes A. (2007).
Interruption of CryAB-amyloid oligomer formation by HSP22. J Biol Chem 282, 555-563.
1 Selcen, 0 D., Muntoni, F., Burton, B.K., Pegoraro, E., Sewry, Florence
C.,Manero
Bite, A.V.,
21.59 and
66.41Engel,
(13) A.G.
Read Citations Mutation in BAG3 causes severe dominant childhood muscular dystrophy. Ann
(2009).
Neurol 65, 83-89.
Shen, D., Coleman, J., Chan, E., Nicholson, T.P., Dai, L., Sheppard, P.W., and Patton, W.F.
(2011). Novel cell- and tissue-based assays forDiddetecting this researchermisfolded and with
author this publication aggregated
you?
21 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 21
4 12

protein accumulation within aggresomes and inclusion bodies. Cell Biochem Biophys 60,
Emeritus professor
173-185. Add a new Article
Emeritus professor
Simon, S., Dimitrova, V., Gibert, B., Virot, S., Mounier, N., Nivon, M., Kretz-Remy, C., Corset,
V., Mehlen, P., and Arrigo, A.P. (2013). Analysis of the dominant effects mediated by wild
type or R120G mutant of alphaB-crystallin (HspB5) towards Hsp27 (HspB1). PLoS One 8,
e70545.
Overview
Simon, S.,
Contributions
Fontaine, Info
Timeline
J.M., Martin,

Stats
J.L.,Scores

Sun, X., Hoppe, A.D., Welsh, M.J., Benndorf, R., and
Vicart, P. (2007a). Myopathy-associated alphaB-crystallin mutants: abnormal
phosphorylation, intracellular location, and interactions with other small heat shock
proteins. J Biol Chem 282, 34276-34287. Centre Lon Brard
What are youResidue Simon, working
S., Michiel,on M., Skouri-Panet, F., Lechaire, J.P., Vicart,
R120 is essential for the quaternary structure and
P., and
Apoptosis, CancerTardieu, A. (2007b).
and Development Laboratory
Lyon,functional
France integrity of human
right now? alphaB-crystallin. Biochemistry 46, 9605-9614.
Sridhar, S., Botbol, Y., Macian, F., and Cuervo, A.M. (2012). Autophagy and disease: always
Add your current projecttwo to share
sidesupdates
to aon your work J Pathol 226, 255-273.
problem.
Tam, A.B., Mercado, E.L., Hoffmann, A., and Niwa,About M. (2012). ER stress activates NF-
kappaB by integrating functions of basal IKK activity, IRE1
I have studied Heat and PERK.
shock proteins PLoS
(Hsps), Onethe7,
particularly small
e45078. ones (Hsp27, alphaB-crystallin) and participated in the discovery
Add
Tannous, P., Zhu, H., Johnstone, J.L., Shelton, J.M., Rajasekaran, N.S., Benjamin, I.J.,
Nguyen, L., Gerard, R.D., Levine, B., Rothermel, B.A., and Hill, J.A. (2008). Autophagy is
an adaptive response in desmin-related cardiomyopathy. Proc Natl Acad Sci U S A 105,
9745-9750.
Show your career's best Turner, B.J., Atkin, J.D., Farg, M.A., Zang, D.W., Rembach, A., Lopes, E.C., Patch, J.D., Hill,
A.F., and Cheema, S.S. (2005). Impaired extracellular secretion Add your other ofsocial networksuperoxide
mutant services to Mnd more
the best way to show your peersdismutase
what you've been1working
associates with for
on, create exposure neurotoxicity
your current in familial amyotrophic
updates.
lateral sclerosis. J
projects, and start building yourNeurosci
network. 25, 108-117.
Vabulas, R.M., Raychaudhuri, S., Hayer-Hartl, M., and Hartl, F.U. (2010). Connect Protein
now folding in
the cytoplasm and the heat shock response. Cold Spring Harb Perspect Biol 2, a004390.
Vicart, P., Caron, A., Guicheney, P., Li, Z., Prevost, M.C., Faure, A., Chateau, D., Chapon,
F., Tome, F., Dupret, J.M., Paulin, D., and Fardeau, M. (1998). A missense mutation in the
191 6.16k alphaB-crystallin
12,280 661.97 chaperone 123gene causes a desmin-related myopathy.
Skills and expertise (32) Nat Genet 20, 92-
Publications Reads 95.Citations Impact Points Full-texts View stats
Villella, V.R., Esposito, S., Bruscia, E.M., Maiuri, M.C., Raia, V., Kroemer, G., and Maiuri, L.
(2013). Targeting the Intracellular Environment in Cystic Fibrosis:
6 Cancer Biology 6Restoring Autophagy
Reactive Oxygen Species as
a Novel Strategy to Circumvent the CFTR Defect. Front Pharmacol 4, 1.
Wang, L., Schubert, D., Sawaya, M.R., Eisenberg, D., and Riek, R. (2010). Multidimensional
Article:structure-activity
NF B is a central regulator relationship of a protein in its aggregated states. Angew Chem Int Ed
of protein quality
control Engl 49, 3904-3908.
in response to protein aggregation stresses via Top co-authors View all
autophagy
Wei, modulation
Y. (2014). Autophagic induction of amyotrophic lateral sclerosis-linked Cu/Zn
Carole Kretz 32.81 220.82 (43)
Mathieu superoxide
Nivon Loc Fort dismutase
Pascale Muller 1
Emma
G93A Richetmutant
in NSC34 cells. Neural Regen
Claude Bernard ResLyon
University 9, 116-24.
Wignes, J.A., Goldman, J.W., Weihl, C.C., Bartley, M.G., and Andley, U.P. (2013). p62
expression and autophagy in alphaB-crystallin R120G Chantal mutant Diaz-Latoud
knock-in mouse model of
115,
28.03 126.96 (32)
Source
63 hereditary
0 cataract. Exp Eye Res 263-273. Claude Bernard University Lyon 1
Williams,
Reads CitationsA., Jahreiss, L., Sarkar, S., Saiki, S., Menzies, F.M., Ravikumar, B., and
Rubinsztein, D.C. (2006). Aggregate-prone proteins Stphanie are cleared Simon from25.30 the
70.79 cytosol
(19) by
autophagy: therapeutic implications. Curr Top Dev Biol 76, Universit Paris-Est Crteil Val de Marne ...
89-101.
Article: Meet Our
Wilson, M.I.,Associate
Dooley, Editor:
H.C., and Tooze, S.A. (2014). WIPI2b and Atg16L1: setting the stage
for autophagosome
Andre-Patrick Arrigo formation. Biochem Soc Trans 42, 1327-1334. Ecole Pratique des Hautes Etudes
Wirth, M., Joachim, J., and Tooze, S.A. (2013). Autophagosome formation--the role of ULK1
1 and0 Beclin1-PI3KC3 complexes in setting the stage. Semin Florence Manero Biol
Cancer 23,
21.59 66.41 (13)
301-309.
ReadZanotto-Filho,
Citations A., Braganhol, E., Battastini, A.M., and Moreira, J.C. (2012). Proteasome
inhibitor MG132 induces selective apoptosis in glioblastoma cells through inhibition of
PI3K/Akt and NFkappaB pathways, mitochondrial dysfunction, and activation of p38-
JNK1/2 signaling. Invest New Drugs 30, 2252-2262. Did this researcher author this publication with you?
22 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 22
4 12

Emeritus professor

Overview
Info
Stats
Scores
Centre Lon Brard

Lyon, France
right now?
Add

Connect now


Figure 1: Amino acid analog, MG132 treatments and overexpression
Carole Kretz 32.81 of mutated
220.82 (43) forms of
Stphanie
HspB5 Simonand Baptiste
SOD1Guey Malenn
induce Fournier
the Andr-
formation of protein aggregates. HeLa cells were either
Chantal Diaz-Latoud
untreated (NT) or submitted to 5 and 15 mM azetidine (A) or28.03
canavanine
126.96 (32)(B) treatments or to
Source
Reads
2.5 and 5 M MG132 treatments (C) during 6 hours and allowed to recover in fresh culture
Citations
medium for 16h. HeLa cells were transiently transfected Stphanie Simon 25.30 70.79 (19)
with control (Cont) plasmids (see
materials
Article: Meet Ourand methods)
Associate Editor: or plasmids expressing wild type HspB5 or its mutated form
Andre-Patrick Arrigo Ecole Pratique
HspB5R120G (D) or wild type SOD1-EGFP or its mutated formsdesSOD1G93A-EGFP
Hautes Etudes and
1 SOD1G85R-EGFP
0 (E) and were analyzed 48h after Florence
transfection. Cells
Manero 21.59 66.41were
(13) fixed,
Read Citations
permeabilized and stained with antibodies against Multi-Ubiquitin (red staining), pericentrin
(indicator of MTOC localization, green staining) (A, B and C) or HspB5 (red staining) and
pericentrin
Chapter: Immense (green staining) Associated
Cellular Implications (D). Nuclei
to were stained
Did with Hoechst
this researcher author(blue) and with
this publication cells
you? were
23 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 23
4 12

Emeritus professor

Overview
Info
Stats
Scores
Centre Lon Brard

Lyon, France
right now?
Add

Connect now


Carole Kretz 32.81 220.82 (43)
Chantal Diaz-Latoud
28.03 126.96 (32)
Source
Reads Citations

1 0 Florence Manero 21.59 66.41 (13)

Read Citations
H...
Source
Page 24
Skip No Yes
Questions
Topics (7)
4 12

Emeritus professor

Overview
Info
Stats
Scores
Centre Lon Brard

Lyon, France
right now?
Add

Connect now


Carole Kretz 32.81 220.82 (43)
Chantal Diaz-Latoud
28.03 126.96 (32)
Source
Figure
Reads 2:
Citations Protein aggregation stresses stimulate NFB activity. pNFBLuc stable HeLa
transformants were either left untreated (NT) or treated with 5 orParis-Est
Universit 15 mM Crteilazetidine (Aze) or
Val de Marne ...
Article: Meet Our Associate

canavanine (Cana)Editor:
(A) or with increasing concentrations (0.5 to 10
Catherine PaulM) of MG132
30.45 (B) during
151.49 (16)
6 hours. As a positive control for NFB induction, cells were also submitted to a 90 min heat
1 shock
0 treatment (HS) at 43C (A). In panels C and D, HeLa
Florence Manero 21.59 66.41 (13)
cells ofwere
University Angerstransiently transfected
Read Citations
or not (NT) with control plasmids (Cont) or plasmids expressing wild type HspB5 or its
mutated form HspB5R120G (C) or wild type SOD1-EGFP or its mutated forms SOD1G93A-
25
Neurodegeneration in Alzheimers, Parkinsons and
Benjamin Gibert
H...
Source
Skip No Yes
Questions
Topics (7)
Page 25
4 12

EGFP and SOD1G85R-EGFP (D). (E) pNFBLuc HeLa cells were either transfected with an
Emeritus professor
empty
Emeritus vector
professor Add mutants
or with expression vectors for dominant negative a new of
Article
IB (pLXSN-
IB) or of IKK (pRK5-IKK(K44A)). The day after, cells were treated with 2000 U/ml
TNF during 4 hours followed by 16h of recovery or submitted to the same treatments as
Overview
Contributions
describedTimeline
above
Info and Cana

(Aze
Stats 15 Scores
mM, MG132 5 M or expression of wild type and

mutated forms of HspB5 and SOD1). Inserts: immunoblot analysis of HspB5 and SOD1-
EGFP levels in transfected HeLa cells, 48 hours after transfection. Luciferase activity was
Centre Lon Brard
measured 16 hours after the treatments or 48h after the transfections.
Lyon, France Results are presented
right now?as fold stimulation (ratio between luciferase activity after treatments vs luciferase activity in
Add your current project to share updates
untreated on your work
(NT) counterparts). Results are presented as means SD, n3.
Add

Connect now


Carole Kretz 32.81 220.82 (43)
Chantal Diaz-Latoud
28.03 126.96 (32)
Source
Reads Citations

1 0 Florence Manero 21.59 66.41 (13)

Read Citations
26 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 26
4 12

Emeritus professor

Overview
Info
Stats
Scores
Centre Lon Brard

Lyon, France
right now?
Add

Connect now


Carole Kretz 32.81 220.82 (43)
Chantal Diaz-Latoud
28.03 126.96 (32)
Source
Reads Citations

1 0 Florence Manero 21.59 66.41 (13)

Read Citations
Figure 3: Amino acid analog, MG132 treatments and overexpression of mutated forms of
HspB5 and SOD1 increase autophagic flux. HeLa cells were either non-treated (NT) or
submitted
Small to azetidine
Stress Proteins Expression:or canavanine
Impacts on Human(5 or 15 mM) treatments (A) or to MG132 (0.5 to 5 M)
27 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 27
4 12

treatments (B) during 6 hours. C) and D) Cells were left untreated (NT) or transiently
Emeritus professor
transfected
Emeritus professor Add awild
with control plasmids (Cont) or plasmids expressing new typeArticle
HspB5 and its
mutated form HspB5R120G (C) or wild type SOD1-EGFP and its mutated forms SOD1G93A-
EGFP and SOD1G85R-EGFP (D). Panels E to G, HeLa cells were submitted to the same
Overview treatments
Contributions as described
Timeline Info above

Statsin the Scores

presence of 5 M of cathepsin inhibitors pepstatin A

(PepA) and E64d. The inhibitors were added concomitantly to the treatments or just after
transfections and were not removed until protein extraction. Total protein extracts were
Centre Lon Brard
prepared 16 hourson Apoptosis, Cancer and Development Laboratory
after treatments and 24 hours after transfections and submitted to SDS-
Lyon, France
right now?Page. Immunoblots were probed with antibodies against LC3-I and -II, HspB5, EGFP and
Actin
Add your current project (as a
to share loading
updates control).
on your work The histograms in every panel show LC3-II/Actin ratios. LC3-
and get noticed for your research. About and Actin bands of western blots
II/Actin ratios were calculated from quantifications of LC3-II
by Image J (n=3, ratio was set at 1.0 for NT or ones
Cont transfected cells). Pictures are
(Hsp27, alphaB-crystallin) and participated in the discovery
Add
representative of 3 independent experiments.

Connect now


Carole Kretz 32.81 220.82 (43)
Chantal Diaz-Latoud
28.03 126.96 (32)
Source
Reads Citations

1 0 Florence Manero 21.59 66.41 (13)

Read Citations
28 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 28
4 12

Emeritus professor

Overview
Info
Stats
Scores
Centre Lon Brard

Lyon, France
right now?
Add

Connect now

Figure 4: p65-depletion alters autophagy induction by protein aggregation stresses. Control
(HeLa) and p65-depleted cell lines (p65-KD#2) were either left untreated or submitted to
amino acid analog (azetidine and canavanine) (A) or MG132 treatments (B) as described in
Figure
control 3. Panels
in response C aggregation
to protein and D, cell lines
stresses via were untreated (NT) or transiently transfected withView all
Top co-authors
control plasmids (Cont) or plasmid expressing wild type orCarole
mutated
Kretz forms of HspB5
32.81 220.82 (43) (C) and
Stphanie Simon Baptiste(D),
SOD1-EGFP Guey Malenn Fournier Andr-
as described in Figure 3. Total protein extracts were analyzed in
Chantal Diaz-Latoud
immunoblots probed with LC3, HSPB5, EGFP and Actin antibodies. The
28.03 126.96 (32)histograms in every
Source
panel show LC3-II/Actin ratios that were calculated as described in Figure 3. Pictures are
Reads Citations
representative of 4 independent experiments. Universit Paris-Est Crteil Val de Marne ...

1 0 Florence Manero 21.59 66.41 (13)

Read Citations
29 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 29
4 12

Emeritus professor

Overview
Info
Stats
Scores
Centre Lon Brard

Lyon, France
right now?
Add

Connect now


Carole Kretz 32.81 220.82 (43)
Chantal Diaz-Latoud
28.03 126.96 (32)
Source
63 0
Figure 5: p65-deficient cells show increased protein aggregation in response to various
Reads Citations
protein aggregation stresses. Control (HeLa) and p65-depleted cell lines (p65-KD#2) were
either
Article: Meetuntreated (NT)
Our Associate or
Editor: treated during 6 hours with 15 mM azetidine (Aze) or canavanine
Andre-Patrick
(Cana)Arrigo
(A) Ecole Pratique
or with 2.5 and 5 M of MG132 (B). Panels C and D, celldeslines
Hauteswere
Etudes untreated or
1 transiently
0 transfected with plasmids expressing HspB5wt and HspB5R120G
Florence Manero 21.59 (C)
66.41or SOD1wt-
(13)
Read Citations
EGFP, SOD1G93A-EGFP and SOD1G85R-EGFP (D). Immunofluorescence analyses were
performed 16 hours after the treatments or 48 hours after transfections. Cells were fixed,
permeabilized
Chapter: andImplications
Immense Cellular stained with Multi-Ubiquitin
Associated to (red staining) and Pericentrin
Did this researcher (green
author this publication with staining)
you?
30 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 30
4 12

antibodies (A and B) or with HspB5 antibody (red staining) (C). Fluorescent antibodies,
Emeritus professor
SOD1-EGFP
proteins and nuclei stained with Hoechst (blue staining) were Article
visualized under
fluorescence microscope. In each panel, the percentage of cells containing aggregates with a
larger or smaller than 3 m was determined and showed in histograms (mean SD), n=3.
Overview
Info
Stats
Scores
Centre Lon Brard

Lyon, France
right now?
Add

Connect now


Carole Kretz 32.81 220.82 (43)
Chantal Diaz-Latoud
28.03 126.96 (32)
Source
Reads Citations

1 0 Florence Manero 21.59 66.41 (13)

Read Citations
31 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 31
4 12

Emeritus professor

Overview
Info
Stats
Scores
Centre Lon Brard

Lyon, France
right now?
Add

Connect now


Carole Kretz 32.81 220.82 (43)
Chantal Diaz-Latoud
28.03 126.96 (32)
Source
Reads Citations
Figure 6: NFB upregulates BAG3 and HspB8 expressionStphanie
after Simon 25.30
various 70.79aggregation
protein (19)
stresses.
Article: Meet Our(A) Control
Associate (HeLa)
Editor: and p65-depleted (p65-KD#2) cell lines were either untreated
Andre-Patrick
(NT) orArrigo
treated with 5 and 15 mM azetidine (Aze) and Ecole Pratique des Hautes
canavanine (Cana)Etudes
or with 5 M
1 MG132,
0 during 6 hours. 16 hours after treatments, total protein extracts
Florence Manero were
21.59 prepared
66.41 (13) and
Read Citations
analyzed by immunoblots probed with antibodies against BAG3, HspB8 and Actin (as a
loading control). B) Same analysis as in A) but with HeLa and p65-KD#2 cell lines transiently
transfected
Chapter: with control
Immense Cellular plasmids
Implications (Cont)
Associated to or plasmids
Did expressing HspB5wt,
this researcher author HspB5R120G,
this publication with you?
32 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 32
4 12

SOD1wt-EGFP, SOD1G93A-EGFP or SOD1G85R-EGFP. Analysis was performed 35h post-
Emeritus professor
transfection.
Emeritus professor Add aby
Expression of HspB5 and SOD1-EGFP was checked new Article
probing immunoblots
with anti-HspB5 and anti-EGFP antibodies. BAG3 and HspB8 levels were quantified; the
densitometric measurements were normalized to the corresponding Actin bands and ratios
Overview were
Contributions calculated
Timeline between
Info HeLa

Statsor p65-KD#2-treated
Scores
cells vs non treated counterparts and

reported in graphs in each panel (n=3).
Centre Lon Brard

Lyon, France
right now?
Add

Connect now


Carole Kretz 32.81 220.82 (43)
Chantal Diaz-Latoud
28.03 126.96 (32)
Source
Reads Citations

1 0 Florence Manero 21.59 66.41 (13)

Read Citations
33 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 33
4 12

Emeritus professor

Overview
Info
Stats
Scores
Centre Lon Brard

Lyon, France
right now?
Add

Connect now


Carole Kretz 32.81 220.82 (43)
Chantal Diaz-Latoud
28.03 126.96 (32)
Source
Reads Citations

1 0 Florence Manero 21.59 66.41 (13)

Read Citations
Figure 7: BAG3 co-localizes with protein aggregates induced by various aggregation

stresses. (A to D) HeLa cells were either non-treated (A) or treated with 15 mM azetidine (B)
and
Small canavanine
Stress (C) or with
Proteins Expression: 5 M
Impacts MG132
on Human (D) during 6 hours. 16 hours after treatments, cells
34 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 34
4 12

were fixed and analyzed by confocal microscopy for BAG3 and Multi-ubiquitine localization.
Emeritus professor
(E professor
Emeritus to I) Cells were transiently transfected with plasmidsAdd a new HspB5wt
expressing Article (E) or
HspB5R120G (F) or SOD1 wt-EGFP (G), SOD1G85R-EGFP (H) and SOD1G93A-EGFP (I).
48 hours after transfection, cells were fixed and analyzed by confocal microscopy for BAG3,
Overview HspB5
Contributions and SOD1-EGFP
Timeline Info localization.

Stats Single channels and channels overlay (merge) are

Scores

shown. The intensity of red and green fluorescence measured along portions (white lines in
inserted pictures) that crossover aggregates are plotted for each condition. Each table
Centre Lon Brard
indicates on
the r2 and Apoptosis, Cancer and Development
the percentage of aggregates co-localizing with BAG3. r 2 are theLaboratory
means
Lyon, France
right now?of the squares of Pearsons correlation coefficient (mean SD) in aggregate-portions and in
random
Add your current project aggregate-devoid
to share updates on your work portions of the cells; a value close to 0 indicates a lack of co-
and get noticed for your research. About co-localization, because all values
localization, whereas a value close to 1 indicates complete
were positive. The r2 values of the aggregates-containing portions versus aggregates-devoid
Add
portions are statistically different with a p-value <0.001. Percentage of co-localization of
BAG3 with multi-ubiquitinated aggregates (mean SD) is also indicated in tables and was
determined by geometrical center based co-localization. Pictures are representative of 3
identical experiments. Add your other social network services to Mnd more
Connect now


Carole Kretz 32.81 220.82 (43)
Chantal Diaz-Latoud
28.03 126.96 (32)
Source
Reads Citations

1 0 Florence Manero 21.59 66.41 (13)

Read Citations
35 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 35
4 12

Emeritus professor

Overview
Info
Stats
Scores
Centre Lon Brard

Lyon, France
right now?
Add

Connect now


Carole Kretz 32.81 220.82 (43)
Chantal Diaz-Latoud
28.03 126.96 (32)
Source
Reads Citations

1 0 Florence Manero 21.59 66.41 (13)

Read Citations
Figure 8: HspB8 co-localizes with protein aggregates induced by various aggregation

stresses. Same as in figure 7, but cells were analyzed by confocal microscopy for muti-
ubiquitin,
Small HspB5,
Stress Proteins SOD1-EGFP
Expression: and
Impacts on HspB8
Human localization.
36 and
H...
Source
Skip No Yes
Questions
Topics (7)
Page 36
4 12

Emeritus professor

Overview
Info
Stats
Scores
Centre Lon Brard

Lyon, France
right now?
Add

Connect now


Carole Kretz 32.81 220.82 (43)
Chantal Diaz-Latoud
28.03 126.96 (32)
Source
Reads Citations

1 0 Florence Manero 21.59 66.41 (13)

Read Citations
H...
Source
Skip No Yes
Questions
Topics (7)

tmpF02E TMP

Enviado por

Dados do documento

Título original

Direitos autorais

Formatos disponíveis

Compartilhar este documento

Compartilhar ou incorporar documento

Opções de compartilhamento

Você considera este documento útil?

Este conteúdo é inapropriado?

Direitos autorais:

Formatos disponíveis

tmpF02E TMP

Enviado por

Direitos autorais:

Formatos disponíveis

4 12

Article Apr 2016 Molecular biology of the cell

Andre Patrick Arrigo 42.20

Emeritus professor Add a new Article

aggregation stresses via autophagy modulation

Mathieu Nivon*, Loc Fort*, Pascale Muller*, Emma Richet* ||

UMR5310, INSERM U1217, Institut NeuroMyoGne.

Article: NF B is a central regulator of protein quality

Questions Supplemental Material can be found at: 1

Andre Patrick Arrigo 42.20

Scoresor familial amyotrophic lateral sclerosis. To fight

Andre Patrick Arrigo 42.20

Andre Patrick Arrigo 42.20

In this study, we show that NFB-depleted cells are unable

Patrick Arrigo Claudio Hetz Julie D Atkin Carole Kretz-Remy

1 0 Florence Manero 21.59 66.41 (13)

Andre Patrick Arrigo 42.20

Patrick Arrigo Claudio Hetz Julie D Atkin Carole Kretz-Remy

Andre Patrick Arrigo 42.20

Andre Patrick Arrigo 42.20

Andre Patrick Arrigo 42.20

2009). After azetidine, canavanine (Figure 4A)

Andre Patrick Arrigo 42.20

Patrick Arrigo Claudio Hetz Julie D Atkin Carole Kretz-Remy

Andre Patrick Arrigo 42.20

azetidine, canavanine and MG132 treatments (Figure 7 and 8, A to D). We observed a

Andre Patrick Arrigo 42.20

Andre Patrick Arrigo 42.20

Scores proteins accumulate and are usually gathered

Andre Patrick Arrigo 42.20

Scores of protein aggregates (Martinez-Vicente and

Andre Patrick Arrigo 42.20

To determine how NFB could modulate autophagy in response to proteotoxicity, we focused

Andre Patrick Arrigo 42.20

indicating that the NFB-dependent upregulation of6BAG3

Andre Patrick Arrigo 42.20

MATERIALS AND METHODS

E64d, pepstatin A, L-azetidine-2-carboxylic acid, L-canavanine sulfate, Hoechst 33258 and

Andre Patrick Arrigo 42.20

Andre Patrick Arrigo 42.20

Scores axiovert 200M or a Zeiss Axio Imager Z1

6 Cancer Biology 6 Reactive Oxygen Species

Andre Patrick Arrigo 42.20

associated athanogene 3 interactome analysis reveals colleagues

Dahlmann, B. (2007). Role of proteasomes in disease.6BMC Biochem

Andre Patrick Arrigo 42.20

Andre Patrick Arrigo 42.20

Andre Patrick Arrigo 42.20

Andre Patrick Arrigo 42.20

Emeritus professor Add a new Article

Centre Lon Brard

Connect with more colleagues

191 6.16k 12,280 661.97 123 Skills and expertise (32)

6 Cancer Biology 6 Reactive Oxygen Species

Article: NF B is a central regulator of protein quality

Andre Patrick Arrigo 42.20

Emeritus professor Add a new Article

Centre Lon Brard

Connect with more colleagues

191 6.16k 12,280 661.97 123 Skills and expertise (32)

6 Cancer Biology 6 Reactive Oxygen Species

Article: NF B is a central regulator of protein quality

Article: Meet Our Associate Editor:

Mathieu Nivon, Loc Fort, Pascale Muller, Emma Richet ||