# IN VITRO REPLICATION OF POLIOVIRUS

> **NIH NIH R01** · UNIVERSITY OF CALIFORNIA LOS ANGELES · 1993 · $194,849

## Abstract

The long term objective of this proposal is to determine the mechanism by
which the single-stranded RNA genome of poliovirus, the prototype of
prototype of picornaviruses, is replicated in infected cells.  The
hypothesis that poliovirus genome-linked protein, VPg (or a precursor to
VPg, Pre-VPg) is used as a primer for initiating synthesis of poliovirus
plus- and minus-strand RNAs will be tested.  The virus-specific proteins
3Dpol (RNA-dependent RNA polymerase) and Pre-VPg and a host a cell protein
(host factor, HF) involved in viral replication will be purified using
various chromatographic techniques.  Purified proteins and appropriate
serological reagents will be used to study the mechanism of viral RNA
replication in vitro.  Specifically, the requirements for uridylylation of
Pre-VPg will be examined using biochemical and serological techniques.
Whether uridylylated Pre-VPg (Pre-Vpg-pU or Pre-VPg-pUpU) can be used as a
protein-nucleotide primer for in vitro synthesis of poliovirus plus- and
minus-strand RNAs will be determined.  The protein-protein interaction of
3Dpol,HF and Pre-VPg will be studied using biochemical and serological
techniques.  The mechanism of viral RNA replication will be further studied
using mutant polioviruses defective in RNA synthesis.

The normal function of host factor in uninfected cells will be examined.
Since a double-stranded RNA (dsRNA)-activated protein kinase activity which
phosphorylates the alpha-subunit of eukaryotic initiation factor-2 (eIF-2)
copurifies with host factor, the exact relationship between host factor and
dsRNA-activated kinase will be examined using biochemical and serological
techniques.  Furthermore, the mechanism by which poliovirus prevents
increased phosphorylation of eIF02lalpha despite activation of dsRNA-
activated kinase in infected cells, will be examined.

Elucidation of the mechanism of replication of poliovirus will certainly
fill the remaining large gap in knowledge concerning replication of this
medically important virus group which includes those inducing common cold,
infectious hepatitis, encephalomyocarditis and foot-to-mouth disease in
human and animals.

## Key facts

- **NIH application ID:** 3127796
- **Project number:** 5R01AI018272-12
- **Recipient organization:** UNIVERSITY OF CALIFORNIA LOS ANGELES
- **SAM.gov UEI:** RN64EPNH8JC6
- **Principal Investigator:** ASIM DASGUPTA
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1993
- **Award amount:** $194,849
- **Award type:** 5
- **Project period:** 1981-09-30 → 1995-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3127796

## Citation

> US National Institutes of Health, RePORTER application 3127796, IN VITRO REPLICATION OF POLIOVIRUS (5R01AI018272-12). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3127796. Licensed CC0.

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