BACTERIOPHAGES and their life cycle

Saumil S. Sharma
What do you know about
Bacteriophages?
 Introduction
• A bacteriophage (informally, phage) is a virus that infects
and replicates within bacteria.
• The term is derived from 'bacteria' and the Greek
phagein "to devour / to eat up".
• Bacteriophages are composed of proteins that encapsulate
a DNA or RNA genome, and may have relatively simple or
elaborate structures. Their genomes may encode as few as
four genes, and as many as hundreds of genes.
• Phage replicate within bacteria following the injection of
their genome into the cytoplasm.
 Introduction
• Definition: Phages are obligate intracellular parasites that
multiply inside bacteria by making use of some or all of the
host biosynthetic machinery.
• Felix d'Herelle coined the term “Bacteriophage”.
Bacteriophage means to eat bacteria, and are called so
because virulent bacteriophage can cause the compete
lysis of a susceptible bacterial culture.
• They occur widely in nature and can readily be isolated
from feces and sewage. There are at least 12 distinct
groups of bacteriophages, which are very diverse
structurally and genetically.
 Discovery
• Bacteriophages were discovered more than a century ago. In
1896, Ernest Hanbury Hankin, a British bacteriologist (1865 –
1939), reported that something in the waters of rivers in India
had unexpected antibacterial properties against cholera and this
water could pass through a very fine porcelain filter and keep
this distinctive feature (Hankin, 1896).
• In 1915, the British bacteriologist Frederick Twort (1877–1950),
Superintendent of the Brown Institution of London, discovered a
small agent that killed colonies of bacteria in growing cultures.
• Felix d'Herelle (1873-1949) discovered the agent killing bacteria
independently at the Pasteur Institute in France in 1917. He
bserved that cultures of the dysentery bacteria disappear with
the addition of a bacteria-free filtrate obtained from sewage.
 Composition
• Depending upon the phage, the nucleic acid can be
either DNA or RNA but not both. The nucleic acids of
phages often contain unusual or modified bases,
which protect phage nucleic acid from nucleases that
break down host nucleic acids during phage infection.
Simple phages may have only 3-5 genes while complex
phages may have over 100 genes. Certain phages are
known have single stranded DNA as their nucleic acid.
• Protein
– Protection & Infection
Morphology
 Morphology
• The phages have a simple structure comprising of a genome which is
surrounded by a protein covering known as phage capsid. The genome
usually comprises of single, linear, double stranded DNA molecule.
• Large phages generally consist of a head and a tail.
• The Head is a hexagonal structure which encompasses the genome.
The head consists of tightly packed core of nucleic acid surrounded by
a protein coat or capsid and measure in size from 28-100 nm.
• The Tail is a cylinderical structure which is used as an organ of
attachment as well as the conduit through which phage DNA passes
into the host cell. It consists of a hollow core and is surrounded by a
contractile sheath and a terminal base plate to which are attached
pins or tail fibres or both.
 Morphology
• Most phages range in size from 24-200 nm in length. T4 is among the
largest phages; it is approximately 200 nm long and 80-100 nm wide.
All phages contain a head structure, which can vary in size and shape.
Some are icosahedral (20 sides) others are filamentous.
• The head encloses nucleic acid and acts as the protective covering.
Some phages have tails attached to the phage head.
• The tail is a hollow tube through which the nucleic acid passes during
infection. T4 tail is surrounded by a contractile sheath, which contracts
during infection of the bacterium. At the end of the tail, phages like T4
have a base plate and one or more tail fibers attached to it.
• The base plate and tail fibers are involved in the binding of the hage
to the bacterial cell. Not all phages have base plates and tail fibers.
 Types (morphology)
• T-even phages such as T2,
T4 and T6 that infect E.coli
• Temperate phages such as
lambda and mu
• Spherical phages with
single stranded DNA such
as PhiX174
• Filamentous phages with
single stranded DNA such
as M13
• RNA phages such as Qbeta
 Types (life cycle)
A phage can either be virulent or temperate
• Virulent phages: lyse or kill their hosts after
infection (lytic)
• Tempererate phages:
can achieve a state where their genome
replicates along with the host genome without
killing it (lysogenic)
 Life Cycle
Phages exhibit two different types of life cycles:
• Lytic cycle in which intracellular multiplication of
the phage results in the lysis of the host bacterium
and release of progeny virions, also known as
virulent cycle.
• Lysogenic cycle in which phage DNA becomes
integrated into the bacterial genome and
replicates with the bacteria without causing any
harm to the host cell, also called as temperate cycle.
 Terms
• Lyticgrowth: you have replication of phage DNA and synthesis
of new coat proteins. They combine to form new page
particles that are released by lysis of host cell
• Lysogenicgrowth: Involves integration of phage DNA into the
chromosome of the host cell where it replicates during cell
division
• Prophage: integrated phage
• Lysogen: bacterium harbouring the prophage
• Lysogenic bacteria has immunity against further infection
from a similar bacteriophage
• Induction: switch from lysogenic to lytic pathway
 Lytic Cycle
• The lytic cycle of bacteriophages can be described in following steps:
• Adsorption is a very rapid process and is completed within minutes.
• Penetration follows adsorption. Phages inject the phage DNA into the periplasmic
membrane space between the cell wall and the cell membrane. In most phages the empty
head and tail remains outside the bacterium after the injection of phage DNA. After entry,
phage DNA converts into circular form and the process is called as circulation of phage DNA.
• Eclipse Phase, for several minutes following infection active phage is not detectable. During
this period a number of new proteins are being synthesized which include enzymes necessary
for synthesis of phage DNA. After the eclipse phase is over, the synthesis of bacterial proteins
stops and that of protein subunits of phage's head and tail starts.
• Assembly, The protein subunits of the phage head and tail aggregate spontaneously (self-
assembly) to form the compact capsid.
• Maturation, The assembly of the phage components into the mature infective phage
particle is known as maturation.
• Lysis and Liberation of New Phages
The interval between the infection of a bacterial cell and the first release of infectious phage
particles is known as latent period. The average yield of progeny phages from the infected
bacterial cells is known as burst size. The period during which infectious phages released rise
is known as the rise period.
 Kinetics of phage infection
 0 min. Attachment of T2 to a susceptible E. coli cell
 1 min. Inject DNA into cell
 1-7 min. Transcribe and translate early genes
 block bacterial DNA synthesis and degrade host chromosomal DNA
 block transcription of host mRNAs
 block translation of host proteins
 small amounts of early proteins produced (catalytic functions)
 transcription from single phage genome
 7-15 min. Replication of phage DNA
 10-20 min. Translation of phage late proteins (structural)
 transcribed from new phage DNA (many copies of template)
 need large amounts of these proteins to build new virions
 18-25 min. Assembly of new phage particles (end of eclipse
period)
 25 min. Lysis of host cell and release of progeny (end of latent
period)
 Lysogenic Cycle
• Infection with every phage does not result into lysis of the host
cell. Some phages on entering into the cell, do not multiply in the
manner described above for virulent phages. Instead, these
integrate into the genome of the host chromosomes. The
integrated genome of the phage is called as prophage. This
phenomenon is known as lysogeny. Bacteria which have prophage
integrated into their chromosome are designated as lysogens and
such phages are called as lysogenic or temperate phages.
• Lysogeny - After entering the host cell, the DNA gets integrated
into the host DNA. The host cell, now called a lysogenic cell,
replicates the phage DNA (prophage) every time it divides.
Infection by a temperate phage can also lead directly to a lytic
cycle, and on rare occasions, a prophage excises from the bacterial
chromosome and initiates a lytic cycle.
 Importance of Bacteriophages
I. Bacteriophages may act as carriers of genes from one bacterium
to another-a process known as transduction. An important
example of this is drug resistance seen in Staphylococcus
aureus.
II. Phage typing is used as an important epidemiological marker
which helps in establishing the path of transmission of
infectious agent and identifying the reservoir of infection.
III. Subtyping of a species or genus is also possible on the basis of
the reaction to phages (vibrios, staphylococci, salmonellae).
IV. Bacteriophages may endow the property of toxigenicity to
some organisms such as C. diphtheriae.
V. Bacteriophages have been extensively utilized in studying host-
parasite relationships.
 Significance of bacteriophages
• Transduction is responsible for transfer of drug resistance, especially in
Staphylococci
• Lysogenic conversion is responsible for acquisition of new characteristics
• Random insertion into bacterial chromosome can induce insertional
mutation
• Epidemiological typing of bacteria (phage typing)
• Lambda phage is a model system for the study of latent infection of
mammalian cells by Retroviruses
• Phages are used extensively in genetic engineering where they serve as
cloning vectors.
• Libraries of genes and monoclonal antibodies are maintained in phages
• They are responsible for natural removal of bacteria from water bodies
 Summary
• A bacteriophage (informally, phage) is a virus
that infects and replicates within bacteria.
• Composed of Head having genetic material
and Tail having proteins
• Two types of life cycles: Lytic and lysogenic
• Useful as vector for genetic engineering
 Question
• What are bateriophages?

• What is the morphology of phages?

 Question
• Elaborate on are two types of life cycles that
bacteriphages show?

• What are the importance of studying

bacteriophages?