BACTERIOPHAGES ( PHAGES)

LEARNING OBJECTIVES

 know the definition of bacteriophages

 know and understand the different life cycles of bacteriophages
 importance of phages in bacterial evolution
 application of phages in research and medicine
 What is a bacteriophage?
 This is a virus that infects bacteria.
…..but are harmless to humans.
 They were discovered in 1915 and 1917 by
Frederick Twort and Felix D’Herelle working
independently.
Phage is from greek, phagein, meaning to eat.
D’herelle coined the term bacteriophage,
meaning, “bacteria eater.”
 are considered to be the most abundant
biological agents on earth.
 can be as many as ten quintillion
 Types of phages

Virtually two types exist:

1. Virulent
2. Temperate
 The difference is due to their lifestyles after infection.
 Virulent phages infect and kill the host (lytic cycle).
 Temperate phages: these infect and stay dormant
until triggered to kill bacterial hosts ( lysogenic and
lytic cycle).
 Characteristics
 their capsules house either DNA or RNA genome.
 to multiply, they get into the host cell, which they
destroy by breaking open to release new viruses.
 some types multiply inside the cells they invade and
escape by lysing the host cell. This interaction is called
productive infection, because more viruses are
produced.
 phages that lyse the cell are termed lytic phages. Lysis (
dissolution).
 lytic phages takeover the host cellular metabolism and
direct it to produce only phages.
Cont…
 Another type of interactive production is when phage
invade the cell, multiply inside, and then leak out or
extrude, without killing the host cell.
 these phages take over only part of the host cell’s
metabolic machinery. The best example is filamentous
phage, M13.
 temperate phage integrate their DNA inside the host
cellular genome or the DNA replicates as a plasmid.
 the phage DNA replicates as the bacterial DNA replicates.
There could be no sign of infection, this is why it is called
latent infection. And the bacterium carrying the phage is a
lysogen and the cell is in lysogenic state.
Cont…
 The phage DNA encodes for proteins which
modify the properties of the host cell, lysogenic
conversion. An example of the temperate phage
that infects E. coli is lambda (λ).
 phage are much more used in experiments than
bacteria. This is because of their simple structure.
More so, they are not a danger to human life.
 The ability of phages to infect and possibly kill
infectious bacterial agents puts forward their
potential as a possible supplement or
replacement for antibiotic agents.
General structure of phage
1. Nucleocapsid: houses the genome of the phage. It is made
up of DNA or RNA enclosed in the capsid. The capsid is itself
made of capsomere protein proteins.
2. The phage collar is a ring-like structure located in the neck
region of a bacteriophage, specifically between the head and
the tail. It plays a crucial role in attaching and organizing
the tail fibers, which are used to attach the virus to a host
bacterial cell. Its also involved in controlling the retraction of
tail fibers.
3. Baseplate has several key functions: host cell recognition,
initiating tail sheath contraction, and triggering genome
ejection.
4. Spikes: attachment to specific receptors, host cell
penetration, enzymatic activity
Lytic Phage Replication by Double-stranded DNA
phages
1. Attachment: The phage binds to specific receptors on the
bacterial cell surface.
2. Entry/Penetration: The phage injects its double-stranded
DNA genome into the host cell's cytoplasm.
3. Replication: The phage DNA is replicated, and phage genes
are transcribed and translated to produce phage proteins.
4. Assembly: New phage particles are assembled from the
replicated DNA and phage proteins.
5. Lysis:The host cell is lysed (burst open), releasing the newly
assembled phage particles to infect other bacteria.
Steps in the replication of T4 phage in E.coli
 Phage Replication in Latent State
 Some phages live in harmony with their host bacteria.
They are called temperate phages.
 90 percent of all phages are temperate.
 The phage DNA is integrated into bacterial chromosome,
where it replicates as the bacterial chromosome
replicates.
 When integrated, the phage DNA is called prophage and
the bacterial cell carrying a prophage is a lysogenic cell or
lysogen.
Phage Replication in a latent state
Roles of phages in microbial genetics
1. As a vector in gene transfer through transduction
 In generalized transduction, random pieces of bacterial DNA are
packed in the phage capsid as the host cell is disintegrated from lysis.
Should the phage inject it in the health host, it may integrate it into its
chromosome altering its genome and that of its generations down the
line.
 In specialized transduction, the phage only transfer a few restricted
genes from the donor cells to recipients. This is carried out only by
temperate phages which undergo a lysogenic life cycle.
Cont…
2. Phage therapy
 A method of delivering virulent phages to a clinically ill patient to
rapidly kill pathogenic bacteria in the area of infection.
- Involves the use of lytic phages, bioengineered phages, or purified
lytic phage proteins.
Advantages of phage therapy
- Targeted therapy: can only bind to specific sites of their host cells
- Low toxicity: they are safe for human use. No side effects.
- Environmental benefits: antibiotics build up in the environs creating
antibiotic resistant microbes.
- Automatic dosing
- Self-amplification
Viroids
 are pathogens much smaller and distinctly different from
viruses.
 mainly consist of a small, single-stranded RNA molecule that
varies in size.
 they have no protein coat and thus resistant to proteases.
 they replicate autonomously within susceptible cells.
 a single viroid RNA molecule is capable of infecting a cell.
 the viroid RNA is circular and resistant to digestion by
nucleases.
 all viroids identified affect only plants and cause serious
diseases.
Cont…
 these diseases include, potato
spindle tuber, citrus exocortis,
cucumber pale fruit disease.

cucumber pale fruit disease

 Lambda DNA integrates into E.coli genome at a specific site through a
process called site-specific recombination.
 Short nucleotide sequences in the DNA of lambda phage and E.coli host
are identical ( homologous), allowing the phage and bacterial DNA to
synapse (pair). In E. coli, this region is between the genes for galactose
metabolism and biotin (vitamin B7) synthesis.
 Phage DNA remain integrated in the bacterial genome and is passed on
to all daughter cells indefinitely. The genes on the integrated DNA must
be silenced/ repressed.
 these genes code for excisase, which is responsible for the removal of
integrated DNA from the host genome.
 one gene in the integrated DNA codes for a repressor that binds to the
viral operator to stop the transcription of excisase. If not, the integrated
DNA is excised and viral proteins are synthesized. Leading to the lytic
phase.