# MOLECULAR BIOLOGY OF POLIOVIRUS RNA REPLICATION

> **NIH NIH R01** · UNIVERSITY OF FLORIDA · 1987 · $117,685

## Abstract

The mechanism of poliovirus RNA replication is being examined in this
study.  The poliovirus RNA dependent RNA polymerase was isolated as a
soluble and template dependent enzyme from the cytoplasm of infected
cells.  Highly purified forms of the polymerase were found to synthesize
full-sized copies of poliovirion RNA other polyadenylated RNAs when
oligo(U) was added as a primer.  In the presence of a cellular protein
component or "host factor", the polymerase will initiate RNA synthesis in
the absence of an oligo(U) primer.  We found that the product RNA that was
synthesized in the presence of the host-factor was about twice the size of
the template RNA.  This and other evidence indicated that the product RNA
was covalently linked to the template RNA.  This suggested that in the
presence of the host-factor the polymerase used a template-priming
mechanism to initiate RNA synthesis in vitro.  We are continuing these
studies to further characterize the in vitro polymerase reaction to
determine if this model is correct.  The activity of the polymerase on
poliovirus minus-strand templates is being examined, and the product RNA
from these reactions is being characterized for its size and polarity.  The
role of VPg in the replication of poliovirus RNA is also being
investigated.  To carry out these studies, VPg was prepared synthetically,
and was used to prepare anti-VPg antibody.  We hope to determine if VPg (or
a VPg precursor protein) plays a role in the initiation of RNA synthesis.
In addition to our studies on the initiation of RNA synthesis, we plan to
further characterize the elongating activity of the polymerase.  The
fidelity of RNA replication in vitro will be measured, and it will be
determined if infectious RNA can be synthesized using minus-strand RNA
templates.  To study viral RNA replication in vivo, we will characterize
the structure of poliovirion RNA isolated from infected cells and will use
a genetic approach to identify and define the functions of viral proteins
required for poliovirus RNA replication.  Poliovirus mutants will be
constructed using infectious DNA plasmids containing the complete viral
sequence.

## Key facts

- **NIH application ID:** 3126231
- **Project number:** 5R01AI015539-09
- **Recipient organization:** UNIVERSITY OF FLORIDA
- **SAM.gov UEI:** NNFQH1JAPEP3
- **Principal Investigator:** James Bert FLANEGAN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1987
- **Award amount:** $117,685
- **Award type:** 5
- **Project period:** 1979-09-01 → 1988-02-29

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3126231

## Citation

> US National Institutes of Health, RePORTER application 3126231, MOLECULAR BIOLOGY OF POLIOVIRUS RNA REPLICATION (5R01AI015539-09). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3126231. Licensed CC0.

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