Poliovirus mutant that contains a cold-sensitive defect in viral RNA synthesis
Population
Monkey kidney cells infected with poliovirus
Design
Preclinical
Authors
Discussion
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Implication
Cold-sensitive 3A mutant blocks early poliovirus RNA synthesis; hypothesis-generating for replication-targeted antivirals or vaccines.
Key Points
- To determine the functional consequences of a single-codon insertion within the 3A region of the poliovirus genome on viral RNA synthesis and replication.
- Engineered a single-codon insertion into the 3A region of an infectious poliovirus cDNA clone.
- Evaluated viral replication, positive- and negative-strand RNA synthesis, and polyprotein processing in monkey kidney cells at permissive temperatures and at 32.5°C.
- Performed temperature-shift experiments to pinpoint the timing of the functional defect during infection.
- The engineered mutant demonstrated a cold-sensitive phenotype with severe depression of both positive- and negative-strand RNA synthesis at 32.5°C.
- Temperature-shift assays revealed that the defective function is required specifically during the early exponential phase of RNA synthesis.
- Polyprotein processing showed incomplete cleavages at the nonpermissive temperature, attributed to the severe reduction in RNA synthesis and virally encoded proteases.
Structured PICO
P
Population
Monkey kidney cells infected with poliovirus
I
Intervention
Single-codon insertion into the 3A region of the viral genome
O
Outcome
Viral RNA synthesis at 32.5 degrees Csurrogate
A single-codon insertion in the 3A region of the poliovirus genome results in a cold-sensitive defect in viral RNA synthesis.
Cite This Study
Berstein et al. (1988) studied this question.