# IN VITRO REPLICATION OF POLIOVIRUS

> **NIH NIH R01** · UNIVERSITY OF CALIFORNIA LOS ANGELES · 1987 · $161,296

## Abstract

The goal of this investigation is to understand the mechanism of
replication of picornaviruses typified by poliovirus.  Proteins involved in
viral RNA replication will be purified using various chromatographic
techniques.  Purified proteins as well as serological reagents will be used
to determine the role of viral and host proteins in the initiation of
poliovirus minus-stranded RNA synthesis.  The hypothesis that poliovirus
genome-linked protein, VPg (or its precursors) is used as a primer for
initiating poliovirus plus- and minus-strand RNA synthesis will be tested
using biochemcal techniques and an antibody to VPg.  Roles of host factor
(HF) and the virus-specific RNA polymerase (P63, replicase) in this
reaction will be examined using biochemical and serological means.
Specific agents, which selectively inhibit initiation of RNA synthesis as
well as phosphorylation of a 63K protein in replicase preparation, will be
used to study the involvement of protein phosphorylation in initiation of
viral RNA replication.  Biochemical and serological techniques will be used
to determine if the kinase activity associated with viral polymerase is
actually due to P63.  Poliovirus ts-mutants that are defective in RNA
synthesis will be used in complementation studies to furter define the
roles of viral proteins in RNA replication.

Molecular organization of the enzymatic machinery responsible for RNA
replication will be studied by biochemical and immunochemical analysis of
the isolated active replicase complex formed by mixing purified viral and
host proteins in vitro.  Also anti-VPg and anti-HF antibodies will be used
to immunoprecipitate proteins complexed with HF and/or VPg from crude,
soluble 7S replicase complex.

Purified proteins will be used to study specific binding of polymerase to
virion RNA using a recently developed RNA binding assay.  Specificity of
transcription of polio RNA (by the replicase) compared to other RNAs will
be determined using biochemical and serological methods.

The long-term goal of this research is to understand the mechanism of
replication of picornaviruses.  Studies will be continued to understand the
mechanism of replication of poliovirus plus-strand RNA synthesis on
minus-strand RNA template.  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 humans and animals.

## Key facts

- **NIH application ID:** 3127792
- **Project number:** 5R01AI018272-07
- **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:** 1987
- **Award amount:** $161,296
- **Award type:** 5
- **Project period:** 1981-09-30 → 1989-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3127792

## Citation

> US National Institutes of Health, RePORTER application 3127792, IN VITRO REPLICATION OF POLIOVIRUS (5R01AI018272-07). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3127792. Licensed CC0.

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