Bacteriophage - Structure, Replication, Uses - Microbe Online

1/5/22, 5:22 PM Bacteriophage: Structure, Replication, Uses • Microbe Online
Bacteriophage: Structure,
Replication, Uses x
Last updated on May 30th, 2021
Bacteriophages “bacteria-eater” are infectious agents that replicate as obligate

intracellular parasites in bacteria with high selectivity. They are the powerful regulators
of bacterial populations in natural ecosystems and are found in the soils, plants, rivers,
and also as the human microbiome.
Phages colonize all body niches, including the skin, oral cavity lungs, gut, and urinary
tract. Although considered sterile, metagenomic analysis has shown that blood contains
bacterial virus of family Myoviridae, Podoviridae, Siphoviridae,
Microviridae, and Inoviridae families.
Phages were first observed in 1915 by Frederic Twort in England and in 1917 by Felix
d’Herelle in France. Felix d’Herelle named them Bacteriophages. In the 1920-1930s,
phages were used to treat bacterial infections in Europe and Soviet Unions but after the
discovery of penicillin use of bacteriophage as antimicrobial agents decreased. Due to
the rise in antimicrobial resistance, there is renewed interest in phage therapy.
Use of phages to treat bacterial or animal infections is called phage therapy.
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Contents x
1 Structure of Bacteriophage x
2 Replication
2.1 Replication of Lytic Phages
2.1.1 Adsorption/Attachment
2.1.2 Penetration
2.1.3 Replication
2.1.4 Assembly
2.1.5 Lysis and Release
2.2 Replication of Temperate Phages
2.2.1 Process of Lysogeny
2.2.2 Lysogenic or Lytic cycle; what determines the fate?
2.2.3 Lysogenic conversion
3 Usefulness of Phages
3.1 Phage Therapy
3.2 Applications in Biotechnology and Research
Structure of Bacteriophage
Schematic representation of main types of phages

(Image source: Brock Biology of Microorganisms)
Bacteriophage structures are diverse, but most of them share some common
characteristics. For example, bacteriophage T4 of Escherichia coli has an icosahedral
head structure made of repeat protein sub-units known as the capsid. This head
structure contains a linear double-stranded viral genome.
Phage genome varies in size from approximately 2 to 200 kilobases per strand of
nucleic acid. Considerable variability is found in the nucleic acid of phages, and it may
be ds DNA, ds RNA, ss DNA, ss RNA. Most known bacteriophages contain dsDNA
genomes.
Structure of a Phage λ (lambda)
The head of bacteriophage T4 is attached to a helical tail through a collar (neck). Tails
contain a series of tail fibers and tail pins at the end. These specialized syringe-like
structures bind to receptors on the cell surface. All bacteriophages do not contain a ‘tail’
structure.
Replication
Phages are classified into two major groups on the basis of their mode of replication.
1. Virulent (lytic phage): Virulent phage replicate in the susceptible bacteria x

producing many copies of themselves and destroys the host cells in the process by
lysis. e.g. T-even; T2 and T4 phages of E. coli. x
2. Temperate phages: Infection by temperate phages can have either of two
outcomes.
Lytic growth or
Lysogeny (non-lytic prophage state)
Replication of Lytic Phages
Replication of lytic bacteriophages
Adsorption/Attachment
Bacteriophage adsorption is the first step to start the infection process. The binding
proteins of the bacteriophage mostly located on the tail fibers interact and recognize
specific receptors present on the bacterial cell wall.
The attachment of phage to its host cell results in morphological changes to both phage
and bacterium facilitating the penetration of the phage into a host bacterium.
Penetration
Lysozyme like enzyme found in the phage tails weakens the bacterial cell wall. Tail
sheath contracts, hollow tube (core) penetrate the weakened cell wall and come in
contact with the cell membrane.
Viral DNA moves from the head via the tube to the bacterial cytoplasm while the phage
capsid remains outside.
Replication x
Phage genes take control of the host cell’s metabolic machinery and directs host cellsxto
produce only viral products. Bacterial DNA is disrupted and their nucleotides are used
as building blocks for new phage.
Phage DNA is transcribed to mRNA using the host cell’s machinery. Translation of mRNA
and capsid proteins and viral enzymes are produced.
Assembly
Head of T4 Phages is assembled in the host cells
cytoplasm from newly synthesized capsid proteins. A viral
dsDNA molecule is packed into each head.
Phage tails are assembled from newly formed base

plates, sheaths, and collars.
When the head is properly packed with DNA, each head is

attached to a tail.
Tail fibers are added and fully formed mature, infective

phages develop. Assembly
Lysis and Release

Lysozyme breaks down bacterial cell wall and bacterial host cells is lysed. Released
phage infects other susceptible bacteria and the new infection process is started.
Burst time: Time from adsorption to cell lysis, generally 20-40 minutes.
Burst size (viral yield): Number of new virions released from each bacterial host
cells. In T4 phage burst size is 50-200.
Replication of Temperate Phages
Temperate phages mostly exhibit lysogeny but can replicate by lytic pathways after x
induction. The best characterized temperate phage is the E. coli phage λ.
x
Lysogeny is a special type of latent viral infection. It’s a stable long term relationship
between the phages and its host in which the phage nucleic acid becomes
incorporated into the host chromosome. The phage genome replicates as a
prophage in the bacterial cell.
In most lysogenic bacteria the genes required for lytic phage development are
repressed and the production of infectious phage does not occur.
Process of Lysogeny
Lambda phages attach to bacterial cells and insert their linear DNA into the bacterial
cytoplasm.
Phage DNA circularize and then integrates into the circular bacterial chromosome at a
specific location. Insertion of a lambda phage into a bacterium alters the genetic
characteristics of the bacterium.
Once established as a prophage, the virus can remain dormant for a long time. Each
time a bacterium divides; the prophage is copied as a part of the bacterial chromosome
and retains in the progeny bacteria. The period of bacterial growth with a prophage x
represents a lysogenic cycle.
x
However, either spontaneously or in response to some outside stimulation, the
prophage can become active and initiate a typical lytic cycle. This process is called
induction.
Lytic and Lysogenic cycle of Bacteriophage
Lysogenic or Lytic cycle; what determines the fate?

The choice depends on the balance between two proteins, the
repressor produced by the c-I gene and x

antagonizer of the repressor produced by the cro gene.
x
If the repressor predominates, transcription of other early genes is shut off and
lysogeny ensues. Transcription is inhibited by binding of the repressor to the two
operator sites that control early protein synthesis.
If the cro gene product prevents the synthesis of sufficient repressor, replication, and
lysis of the cell result.
Lysogenic conversion
Expression of the integrated phage (prophage) genes confers new properties to the
host bacteria. This phenomenon is known as lysogenic conversion. For example;
synthesis of several exotoxins in bacteria, such as diphtheria, botulinum, cholera, and
erythrogenic toxins. Without prophage, these bacteria are non-pathogenic.
Virulence factors encoded by bacteriophages
Organism Virulence factors

Streptococcus Erythrogenic toxin
pyogenes
Escherichia coli Shiga-like toxin

Staphylococcus aureus Enterotoxins A, D, E, Staphylokinase, toxic shock syndrome
toxin-1 (TSST-1)
Clostridium botulinum Neurotoxins C, D, E
Corynebacterium Diphtheria toxin
diphtheriae
Lysogenic conversion is mediated by the transduction of bacterial genes from the donor
bacterium to the recipient bacterium by bacteriophages. During integration into host
genes, the phage loses genes required for replication, this prevents the induction of the
lytic cycle and killing of host bacteria.
Lysogenic conversion
Transduction is the process by which the DNA is mobilized between cell by a virus.
Prophage also provides “immunity” to infection by other phages of the same type. The
gene however does not protect the lysogen against infection by a different type of
temperate phage or by a virulent phage.
Usefulness of Phages
Phages have been studied as model organisms to gain insights into basic genetic
concepts, such as viral gene expression. Researches on bacteriophage gave us much of
our understanding of viruses and many fundamental concepts of molecular biology.
Phage Therapy
Lytic phages can be used as a replacement for antibiotic therapy. Using phages to treat
bacterial infections was developed back in the 1920s and 1930s in Eastern Europe and
the Soviet Union with varying success. Due to the spread of multi-drug resistant
bacteria, phage therapy is re-emerging after a century. In 2019, the FDA approved the
first US clinical trial of intravenously administered bacteriophage therapy.
Applications in Biotechnology and Research
x
Phages are excellent vehicles for horizontal gene transfer by transduction. So, they find
wide use in recombinant-DNA technology to construct mutants and to transfer genes of
interest from one bacterium to another. Phages can also be used as biocontrol agentsx
in agriculture and petroleum industry.
References and further readings

Madigan Michael T, Bender, Kelly S, Buckley, Daniel H, Sattley, W. Matthew, & Stahl,
David A. (2018). Brock Biology of Microorganisms (15th Edition). Pearson.
Manohar, P., Tamhankar, A. J., Leptihn, S., & Ramesh, N. (2019). Pharmacological
and Immunological Aspects of Phage Therapy. Infectious Microbes & Diseases, 1(2),
34–42.
Navarro, F., & Muniesa, M. (2017). Phages in the Human Body. Frontiers in
Microbiology, 8.
Van Belleghem, J. D., Dąbrowska, K., Vaneechoutte, M., Barr, J. J., & Bollyky, P. L.
(2018). Interactions between Bacteriophage, Bacteria, and the Mammalian
Immune System. Viruses, 11(1).
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1/5/22, 5:22 PM Bacteriophage: Structure, Replication, Uses • Microbe Online

Last updated on May 30th, 2021

Bacteriophages “bacteria-eater” are infectious agents that replicate as obligate

Use of phages to treat bacterial or animal infections is called phage therapy.

Schematic representation of main types of phages

Structure of a Phage λ (lambda)

1. Virulent (lytic phage): Virulent phage replicate in the susceptible bacteria x

Replication of Lytic Phages

Replication of lytic bacteriophages

Phage tails are assembled from newly formed base

When the head is properly packed with DNA, each head is

Tail fibers are added and fully formed mature, infective

Lysis and Release

Replication of Temperate Phages

Lytic and Lysogenic cycle of Bacteriophage

Lysogenic or Lytic cycle; what determines the fate?

repressor produced by the c-I gene and x

Virulence factors encoded by bacteriophages

Organism Virulence factors

Escherichia coli Shiga-like toxin

Applications in Biotechnology and Research

References and further readings

Bacteriophage Plaque Assay: Bacterial Typing Methods: Aim, Generalized transduction:

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