Virus Life Cycle Flashcards

1
Q

How do viruses grow?

A
  • inverted microscopes are used to observe virus and cell cultures
  • viruses are NOT visible by light microscopy
  • virus infection may cause cytopathic effects that can be used to study virus replication and infectivity (Not all viruses cause CPE)
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2
Q

Key concepts about viruses

A
  • obligate intracellular parasites
  • no genes for biosynthetic machinery
  • assemble from host cell molecules
  • evolve rapidly compared to host
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3
Q

What does a virus need?

A
  • the right host (tropism)
  • cells with the right receptors (susceptible)
  • appropriate intracellular environment (permissive)
  • biosynthesis machinery
  • abundant building blocks- nucleotides (RNA, DNA), amino acids, ATP, lipids, sugars, etc.
  • time to finish replication
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4
Q

Steps in Virus Replication

A

1) Recognition of the target cell
2) Attachment
3) Entry: Penetration or Fusion
4) Uncoating
5) Transcription of mRNA
6) Protein synthesis
7) Replication of the genome
8) Assembly of virions
9) Egress: lysis, budding, exocytosis

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5
Q

Steps 1 and 2: Recognition and Attachment

A
  • recogntion: interactions between virions and tissues
  • attachment: binding of a virion surface molecule to its specific cellular receptor
  • Entry: Internalization of the virion into the cell. Herpes recognizes the extracellular matrix, then attaches to specific protein receptors
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6
Q

Host Range

A

-the preferred species

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7
Q

Tissue tropism

A

-the preferred cell type

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8
Q

Susceptible

A

-cells that a virus can enter

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9
Q

Permissive

A

-cells that support virus replication and virion synthesis

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10
Q

Abortive infection

A

-replication is incomplete

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11
Q

Step 3: Entry

A
  • virions may use multiple routes to enter cells
  • varies by cell type
  • may have consequences for disease outcome
  • penetration: engulfment of entire virion into cell
  • receptor-mediated endocytosis
  • pinocytosis
  • phagocytosis

-fusion: virion envelop fuses with plasma membrane, leaving parts of the virion behind

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12
Q

Step 4: Uncoating

A
  • release of the genome into the cell
  • for infection to begin, capsids must open to release the genome into the cytoplasm or nucleus
  • uncoating marks the beginning of the “eclipse phase”

Mechanisms of entry and uncoating:

  • at the plasma membrane
  • within endosomes
  • at the nuclear pore
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13
Q

Step 5: Transcription of mRNA

A
  • all viruses must make mRNA
  • viral genome is the template for transcription
  • viral and host transcription factors regulate mRNA synthesis- RNA-dependent RNA polymerase (RDRP)
  • mRNA is made by viral or host polymerases
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14
Q

Step 6: Protein Synthesis

A
  • viral mRNAs are translated into protein by the host machinery- ribosomes, tRNAs, amino acids
  • viral proteins are sorted to site of virion assembly
  • capsid proteins interact with the newly made genomes
  • membrane proteins traffic through the secretory pathway
  • cytosolic proteins accumulate next to the membrane
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15
Q

Step 7: Replication of the Genome

A
  • viral genomes come in many types
  • RNA double or single stranded, + or - sense
  • DNA double or single stranded, + or - sense
  • Linear, circular, segmented, sealed ends, etc
  • polymerases make new genomes using host cell nucleotides
  • viral RNA dependent RNA polymerase (RDRP)
  • viral DNA polymerase
  • host cell DNA polymerase
  • host cell RNA Pol II (RNAP II)
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16
Q

Viral Replication Enzymes

A
  • DNA viruses use viral or host DNA polymerases

- RNA viruses use viral RNA dependent RNA polymerase to make mRNA and genomes

17
Q

Step 8: Assembly of virions

A
  • marks the end of the “eclipse phase”
  • capsid proteins first form an empty shell
  • viral DNA is then inserted into the capsids, making them appear dark on this TEM
18
Q

Capsid assembly

A
  • capsid proteins are usually made late during infection
  • icosahedral and helical capsids self-assemble
  • complex capsids are made of genomes coated with nucleoproteins
  • some capsids mature outside of the cell
19
Q

Virion Envelopment

A
  • enveloped viruses acquire a membrane from a cellular source- ER, Golgi, Plasma membrane
  • viral and cellular proteins are sorted to site of envelopment
  • membrane proteins traffic through the secretory pathway
  • cytosolic proteins accumulate next to the membrane
20
Q

Assembly with Envelopment

A
  • capsid assembly can occur at the same time as envelopment
  • all virion components accumulate at the site of capdi formation, genome incorporation, matrix, glycoproteins and envelopment
21
Q

Step 9: Egress of virions

A
  • virions are released by: budding from the cell, exocytosis, lysis of the cell, cell-to-cell spread
  • virions may transfer to new cells by fusion -> syncytium
22
Q

Routes of virus transfer to new cells: Mechanisms of Egress

A
  • Cell associated virions
  • cell-to-cell spread
  • cell fusion: syncytium formation

-virions released into extracellular space- lysis, budding, exocytosis

23
Q

How a Virus Kills a cell

A

1) Virus enters cell (1 virus)
2) Virus in nucleus, duplicates itself (None, eclipse phase)
3) Cell explodes, releasing thousands of brand-new virus particles (>1000, Burst size)

24
Q

Single-step Virus Growth Curve

A
  • Eclipse: no virus is recovered during the replication and assembly phases
  • maturation and release: virus particles are made and can infect other cells
  • Burst size: the number of infectious viral progeny from a single round of replication
25
Q

Cell lysis produces virus plaques

A
  • virus egress lyses cells
  • lysed cells appear clear when cell monolayers are stained with dye
  • infectious virions are measured in PFUs (Plaque Forming Units)
26
Q

Adenovirus Replication Kinetics

A
  • hours 0-12 hours- eclipse period
  • the intracellular PFU increases from 12 to 28 hours
  • from 0-16 hours is latent phase
  • then after 16 extracellular concentration increases