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Formin a Nuclear Protection
Jorg Renkawitz1 and Michael Sixt1,*
1Institute of Science and Technology Austria (IST Austria), am Campus 1, 3400 Klosterneuburg, Austria
*Correspondence: sixt@ist.ac.at
http://dx.doi.org/10.1016/j.cell.2016.11.024

In this issue of Cell, Skau et al. show that the formin FMN2 organizes a perinuclear actin cytoskeleton that protects the nucleus and its genomic content of migrating cells squeezing through small
spaces.
Cell migration mostly takes place
in three-dimensional microenvironments
composed of cells and extracellular fibers,
where pore diameters are often substantially smaller than the cellular diameter.
Many cells employ pericellular proteolytic
pathways to generate their individual penetrable path. Others can pass through the
interstitium without causing such collateral
damage and are able to undergo considerable deformations in order to squeeze
through narrow gaps. To do so, cells
need to overcome a major hurdle, their
nucleus, which is the largest and stiffest
cellular organelle (McGregor et al., 2016;
Wolf and Friedl, 2011). In this issue, Skau
et al.(2016) report the discovery of a perinuclear actin structure that helps migrating
cells to protect their nucleus content while
migrating through the tissue landscape.
Recent work has found that mechanical
deformation of the nucleus can be a major
threat to genomic integrity of migrating
cells. Challenging the nucleus with strong
mechanical forces, as produced by the
intracellular actin network (Hatch and
Hetzer, 2016) or by migration through
small microenvironmental constrictions
(Raab et al., 2016; Denais et al., 2016;
Irianto et al., 2016), can disrupt the nuclear envelope. This allows nuclear entry
of cytoplasmic factors, which may
together with the concomitant mechanical stress ultimately damage the genome
via hitherto unknown pathways. Nuclear
envelope rupture is buffered by a
shock-absorber role of the intra-nuclear lamin intermediate filament network
(Gruenbaum and Foisner, 2015; Denais
et al., 2016) and the ESCRT pathway
was shown to repair ruptures, thereby
limiting the damage (Raab et al., 2016;
Denais et al., 2016). In the new work
from Skau et al. (2016), the protection
comes from outside the nucleus and is

mediated by the cytoskeletal protein

FMN2.
FMN2 belongs to the formin protein
family, which nucleates and elongates
actin filaments and is also implicated
in adhesion formation and microtubuleactin cytoskeletal crosstalk. By investigating the subcellular localization of
FMN2 in mouse embryonic fibroblasts
(MEFs), the authors identify an unexpected primary localization of FMN2
around the nucleus. Immunostainings
reveal no substantial co-localization with
microtubules and vimentin intermediate
filaments, but rather with actin. Moving
to live-cell microscopy experiments,
the authors confirm and extend these
findings, observing FMN2 together with
actin around the nucleus, specifically at
the trailing half of the nucleus when cells
migrated in 2D and in 3D.
Given this interesting specific localization of FMN2, the authors asked whether
FMN2 was needed for the formation of
this perinuclear actin structure. Indeed,
knockdown of FMN2 results in a loss
of the perinuclear actin bundles in 2D and
3D microenvironments, demonstrating
that FMN2 not only co-localizes with the
perinuclear actin network but is required
for its formation. But what consequence
has the loss of FMN2 and its associated
perinuclear actin network for cell migration? When analyzing cells migrating in
2D, the authors saw an altered nuclear
shape and nuclear positioning but no difference in cell survival compared to control
cells. However, switching to 3D migration
assays, they noted reduced cell viability,
an effect that increases with collagen
concentrationa surrogate of increasing
microenvironmental
complexity
and
decreasing pore sizes.
These results suggest a potential role of
FMN2 in nuclear protection during migra-

1448 Cell 167, December 1, 2016 2016 Elsevier Inc.

tion through microenvironmental pores.

To test this hypothesis, Skau et al.
(2016) first performed transwell migration
assays through differentially sized pores,
resulting in a mild decrease of FMN2
knockdown cells passing through smaller
pores. Second, they analyzed cells
migrating through channels bearing wider
parts and very narrow constrictions.
Whereas control cells largely survived
entering the channels and passing the
constrictions, most FMN2 knockdown
cells already died in the wider part of
the channel with concomitant release of
nuclear material into the cytoplasm. Third
and most explicitly, the authors investigated DNA damage during confined
migration. Using gH2AX as a marker for
DNA double-strand breaks (DSBs), the
authors identified an increased rate of
DSBs in FMN2 knockdown cells, even
more increasing with decreasing pores
sizes in transwell and collagen assays.
Importantly, expression of FMN2-GFP
rescued these phenotypes, whereas a
point mutant deficient only in actin
polymerization (FMN2-I1226A) did not.
Together, these results indicate that
FMN2-induced actin polymerization
protects against DNA damage and increases cellular survival during migration
in confined microenvironments (Figure 1).
Given these results and the high
expression of FMN2 in melanoma, Skau
et al. (2016) investigated the importance
of FMN2 during cancer cell metastasis
using B16-F10 melanoma cells as a
model. They generated a CRISPR-mediated deletion of FMN2, validated the
above-described results in the melanoma
cells, and analyzed the formation of metastases via hematogenic distribution.
Lack of FMN2 strongly diminished the
ability of cells to form metastasis and the
few residual metastases showed a high

thereby decreasing genomic damage

and increasing cellular survival.
Whether cells use alternative strategies
to avoid small pores in order to limit the
events of extreme nuclear deformation
while navigating through the complex
geometry of an interstitium is currently
unknown. Cells might use their mechanosensory machinery to simply circumvent narrow microenvironments or
might switch to proteolytic pathways to
generate a new path. It will also be interesting to understand the contribution
of migration-associated DNA damage
to diseases like the premature aging
associated with nuclear laminopathies or
the malignant transformation of motile
cells.

REFERENCES
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(2016). Science 352, 353358.
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Figure 1. Mechanical Challenge of the Nucleus During Cell Migration and Its Protection
Cells migrating in three-dimensional microenvironments face the challenge of squeezing the nucleus (the
largest cellular organelle) through pores.
(A) Strong mechanical forces such as by migration through small microenvironmental constrictions can
rupture the nuclear envelope and damage the genomic information.
(B) To prevent excessive nuclear damage, the intra-nuclear lamin network acts as a shock absorber, and
the ESCRT pathway repairs nuclear envelope ruptures. Skau et al. (2016) identified a novel additional
mechanism to protect the nucleus, which bases on FMN2-induced perinuclear actin, and increases cell
survival and decreases DNA damage.
(C) The recent identification of considerable nuclear damage and its cellular protection mechanisms opens
a number of important questions to be solved to fully understand how cells cope with the mechanical
challenge the nucleus faces during cell migration.

number of gH2AX foci. This indicated that

the majority of FMN2 knockout cells did
not survive due to high levels of DNA damage during the process of metastasis,
probably due to the missing FMN2 mediated nuclear-protection during confined
cancer cell migration. Thus, FMN2-mediated cellular protection represents a highly relevant cellular mechanism promoting
tumor cell survival.
The identification of an FMN2-based
perinuclear actin network adds to the
recent identification of an Arp2/3-based
(Thiam et al., 2016) and a fascin-based

(Jayo et al., 2016) perinuclear actin

network. Their similarity, their cell type
specificity, and whether they all can serve
a nuclear protective role as described
by Skau et al. (2016) remains to be investigated. Either way, this important newly
identified cytoskeleton-based nucleoprotection mechanism complements the
ESCRT-mediated repair of nuclear envelope rupture (Raab et al., 2016; Denais
et al., 2016) and the shock-absorber role
of the nuclear lamin network (Gruenbaum
and Foisner, 2015) to tolerate nuclear mechanical stress during cellular migration,

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