| Microbiology, 5/e Lansing M Prescott,
Augustana College Donald A Klein,
Colorado State University John P Harley,
Eastern Kentucky University
The Viruses: Bacteriophages
Study Outline- Classification of Bacteriophages
- The most important criteria used for classification are phage morphology and nucleic acid properties
- Most bacteriophages have double-stranded DNA (dsDNA), although single-stranded DNA (ssDNA) and RNA viruses are known
- Most can be placed in one of a few morphological groups: tailless icosahedral, viruses with contractile tails, viruses with noncontractile tails, and filamentous viruses
- Reproduction of Double-Stranded DNA Phages
- Lytic cycle-culminates with the host cell bursting and releasing virions
- The one-step growth experiment
- Reproduction is synchronized so that events during replication can be observed
- Bacteria are infected and then diluted so that the released phages will not immediately find new cells to infect
- The released phages are then enumerated
- Several distinct phases are observed in the viral replication cycle
- Latent period-no release of virions detected; represents the shortest time required for virus reproduction and release; the early part of this period is called the eclipse period, and during this period no infective virions can be found even inside infected cells
- Rise period (burst)-rapid lysis of host cells and release of infective phages; burst size is the number of infective virions released per infected cellc. Plateau period-no further release of infective virions
- Adsorption to the host cell and penetration
- Viruses attach to specific receptor sites (proteins, lipopolysaccharides, teichoic acids, etc.) on the host cell
- Many viruses inject DNA into the host cell, leaving an empty capsid outside
- Synthesis of phage nucleic acids and proteins
- mRNA molecules transcribed early in the infection (early mRNA) are synthesized using host RNA polymerase; early proteins, made at the direction of these mRNA molecules, direct the synthesis of protein factors and enzymes required to take over the host cell
- Transcription of viral genes then follows an orderly sequence due to the modification of the host RNA polymerase and changes in sigma factors
- Later in the infection viral DNA is replicated
- Synthesis of viral DNA sometimes requires the initial synthesis of alternate bases; these are sometimes used to protect the phage DNA from host enzymes (restriction endonucleases) that would otherwise degrade the viral DNA and thereby protect the host
- For some bacteriophages, concatemers of the DNA genome are formed; these are later cleaved during assembly
- The assembly of phage particles
- Late mRNA molecules (those made after viral nucleic acid replication) direct the synthesis of capsid proteins and other proteins involved in assembly (e.g., scaffolding proteins) and release of the virus
- Assembly proceeds sequentially by subassembly lines, which assemble different structural units (e.g., baseplate, tail tube); these are then put together to make the complete virion 3. DNA packaging is still not well understood
- Release of phage particles
- Many phages lyse their host by damaging the cell wall or the cytoplasmic membrane
- A few phages (e.g., filamentous fd phages) are released without lysing the host cell; instead the phages are released through a secretory process
- Reproduction of Single-Stranded DNA phages
- fX174 (+stand DNA virus-virus DNA that has the same sequence as the viral mRNA)
- ssDNA is converted to double-stranded replicative form (RF) by host DNA polymerase
- RF directs synthesis of more RF, RNA and +strand DNA genome
- Filamentous phages (e.g., fd)
- DNA enters via sex pilus
- Replicative form is synthesized
- Replicative form directs mRNA synthesis
- Protein encoded by mRNA then directs phage DNA replication via rolling circle method
- Reproduction of RNA Phages
- Single-stranded RNA phages
- RNA replicase-the virus must provide an enzyme for replicating the RNA genome because the host does not produce an enzyme with this capability
- The RNA genome is usually plus stranded (+) and can act as mRNA to direct the synthesis of the replicase during an initial step after penetration
- +strand RNA is then converted to dsRNA, the replicative form
- Replicative form is then used as a template for production of multiple copies of the genomic (and messenger) +strand RNA
- Capsid proteins are made, and +strand RNA is packaged into new virions
- One or more lysis proteins then function to release the phage
- Only one dsRNA phage has so far been discovered (f6); it infects Pseudomonas phaseolicola and possesses a membranous envelope
- Temperate Bacteriophages and Lysogeny
- Temperate phages are capable of lysogeny, a nonlytic relationship with their hosts (virulent phages lyse their hosts)
- In lysogeny, the viral genome (called a prophage) remains in the host (usually integrated into the host chromosome) but does not kill (lyse) the host cell; the cells are said to be lysogenic (or are called lysogens)
- It may switch to the lytic cycle at some later time; this process is called induction
- Most bacteriophages are temperate; it is thought that being able to lysogenize bacteria is advantageous; supporting this is the observation that certain conditions favor the establishment of lysogeny
- Lysogenic conversion is a change that is induced in the host phenotype by the presence of a prophage and that is not directly related to the completion of the viral life cycle; examples include:
- Modification of lipopolysaccharide structure in infected Salmonella
- Production of diphtheria toxin only by lysogenized strains of Corynebacterium diphtheriae
- Establishment of lysogeny (bacteriophage lambda)
- Two sets of promoters are available to host RNA polymerase
- A repressor protein may be made from genes adjacent to one of these promoters
- If this repressor binds to its target operator before the other promoter is used, then that promoter is blocked and lysogeny is established
- If genes associated with that second promoter are expressed before the repressor can bind to the operator, then the lytic cycle is established
- Induction (the termination of lysogeny and entry into the lytic cycle) will occur if the level of the repressor protein decreases; this is usually in response to environmental damage to the host DNA
- For lambda and most temperate phages, if lysogeny is established, the viral genome integrates into the host chromosome; however, some temperate phages can establish lysogeny without integration
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