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1. Reminder: Actinomycin D is a compound used in the laboratory to inhibit mRNA

ID: 260549 • Letter: 1

Question

1. Reminder: Actinomycin D is a compound used in the laboratory to inhibit mRNA transcription of host cells. (a) If this drug was added to cells infected with a rhabdovirus (e.g., rabies virus), what do you suppose would happen to viral replication? Explain your reasoning. [3 pts.] (b) If this drug was added to cells infected with Influenza virus, what do you suppose would happen to viral replication? Explain your reasoning. [3 pts.] 2. Describe the difference between a protein called a "capsid" compared to a "nucleocapsid." [4 pts.] 3. Consider the information shown in the table below: Influenza A Influenza B Humans only genetic drift only Host range marine mammals Source of genetic variation genetic shift and drift (a) Compare and contrast the terms genetic shift and drift. [3 pts.] (b) Why do Influenza A strains demonstrate both shift and drift while B strains usually experience only genetic drift? [4 pts.] 4. (a) Explain why Influenza viruses were forced to develop the process of "cap-snatching" during their replication cycles? [4 pts.] (b) Pretend that you were working in the research lab that initially discovered the "cap- snatching" mechanism. Describe two experiments or lines of evidence that would have supported this model. Note: I am not asking you to find the original paper(s) and describe what the researchers did. I am asking you to think about what pieces of evidence would have suggested that cap-snatching actually occurred within infected cells. [4 pts.]

Explanation / Answer

1 -

( a )

VSV and rabies virus are only poorly sensitive to cytosine arabinoside at doses as high as 250 ?g/ml which completely inhibit host DNA syntheses.

Rabies virus is more sensitive than VSV to doses of actinomycin D higher than 0.01 gm/ml but this is probably due to the duration of the viral cycle.

( b )

Evidence has been presented to show that actinomycin D inhibits the replication of influenza virus when added during the first 1:30 hours of infection. After 1:30 hours the inhibitory effect progressively decreased.

Actinomycin D has an effect on the replicative form (RF) of the virus.

When the drug was added at 1:30 hours the RF already made disappeared, but when added at 3:30 hours the RF already present remained.

In the absence of the drug, unlabeled uridine added at either time did not chase the uridine-3H out of the RF but, in fact, the total amount of labeled RF increased.

2- Difference – Capsid vs neurocapsid :

The capsid is the protein shell surrounding the genome.

The nucleocapsid is the nucleic acid/protein assembly within the virion.

3 - Compare and contrast : genetic shift and drift :

DRIFT ;

Virus can reolicate in the bird - it can pick up its own genes and never seen the other virsu - Drifts ( happens year to year)

SHIFT :

If they see the other virus - as they are packaged - viruses can pack the other virus gene - Shift ( the big outbreaks - pandemics)

( b )

Drift :

The gradual alteration of the HA and NA within a subype resulting in the reduced ability of antibody to previrous strains to inhibit the HA or N of the mutant virus.

Antigenic drift occurs in both influenza A and B and causes preriodic epidemics.

occurs at a rate of <1% per year at the amino acid level.

Shift :

Appearnce in human population of a new subtype of influenza A virus (HA) and neuraminidase (NA) that is immunologically different from that of isolates circulating previously.

Responsible for world-wide pandemics

Antigenic shift reflects an abrupt change where the new HA may differ by >50% from previously circulating strains.