# MOLECULAR BIOLOGY OF POLIOVIRUS RNA REPLICATION

> **NIH NIH R01** · UNIVERSITY OF FLORIDA · 1989 · $180,451

## Abstract

The molecular mechanisms involved in the replication of poliovirus
RNA will be examined in this study.  We plan to identify viral and
cellular proteins and viral RNA sequences that are important in the
initiation of RNA synthesis, the binding of the polymerase, and the
linkage of VPg to the RNA.  The poliovirus RNA polymerase requires
an oligonucleotide primer or a cellular host factor to initiate RNA
synthesis.  Dimer-sized product RNA is synthesized by the
polymerase in the presence of the host factor.  At least two host
factor activities have been identified.  The host factor associated
with terminal uridylyl transferase activity will be extensively
purified and used to study the initiation of RNA synthesis in
vitro.  Poliovirus cDNA clones will be constructed in transcription
vectors to prepare viral RNA transcripts of different polarities
and containing various deletions and other mutations.  We will
characterize the structure of the product RNAs synthesized by the
polymerase and host factor (TUT, etc.) on both (+) and (-) strand
specific templates and will determine the sequence in the joint
region of dimer-sized product RNAs.  The linkage of VPg to
poliovirus RNA will be studied to determine if this involves a
cleavage reaction and if specific viral RNA sequences are required
for this reaction.  Modified VPg with an altered amino acid
sequence and VPg precursor proteins will be assayed for linkage
activity.  The synthesis of dimer-sized poliovirus RNA in infected
cells will be further characterized.  We will determine if the
synthesis of dimer-sized RNA is increased in cells infected with
certain viral mutants and if guanidine HCl affects the synthesis
of dimer-sized RNA.  The binding of the polymerase to viral RNA
will be examined to determine if high affinity binding sites for
the polymerase exist in (+) and (-) strand viral RNAs.  The optimal
conditions for binding will be determined and other viral and
cellular proteins will be tested for their ability to affect
binding.  A final objective of the study is to quantitate the
polymerase error frequency both in vitro and in vivo.  The
polymerase error frequency will be measured at various sites in
poliovirus RNA under different reaction conditions. The results of
this study should give us a better understanding of the molecular
basis of poliovirus RNA replication.

## Key facts

- **NIH application ID:** 3126232
- **Project number:** 5R01AI015539-11
- **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:** 1989
- **Award amount:** $180,451
- **Award type:** 5
- **Project period:** 1979-09-01 → 1993-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3126232

## Citation

> US National Institutes of Health, RePORTER application 3126232, MOLECULAR BIOLOGY OF POLIOVIRUS RNA REPLICATION (5R01AI015539-11). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3126232. Licensed CC0.

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