# CELLULAR GENE PRODUCT MODULATES FLAVIVIRUS RNA SYNTHESIS

> **NIH NIH R01** · GEORGIA STATE UNIVERSITY · 1991 · $173,103

## Abstract

Experiments are proposed to study the composition and activity of
flavivirus replication complexes using West Nile virus (WNV),
strain E101.  Flaviviruses are small, plus-strand RNA viruses, that
account for a significant amount of human disease worldwide.
After uncoating, the genome RNA, which is the only flavivirus
mRNA, must first be translated to produce the viral replicase
protein(s) needed for viral RNA synthesis.  Currently nothing is
known about the flavivirus replication complex proteins.  Since
two large, viral non-structural proteins, ns5 (96K) and ns3 (67K),
are encoded in the viral genome, it is possible that the plus and
minus strand replication complexes differ.

We propose to identify the WNV replication complex proteins
after extensive purification of the complexes by gradient
centrifugation, column chromatography and immunoprecipitation.
Reconstitution experiments will attempt to demonstrate protein
function.  Host proteins associated with these complexes will also
be studied using cross-linking and RNA binding techniques.  The
involvement of at least one cellular protein in flavivirus RNA
replication is indicated by the existence of a murine flavivirus-
resistance gene.  This resistance is inherited as an autosomal
dominant trait.  Cells from resistant animals synthesize WNV
genomic RNA inefficiently and preferentially amplify viral DI
RNAs.  Comparison of cellular protein components of the WNV
replication complexes obtained from resistant and congenic
susceptible mouse cells may lead to identification of the products
of the alleles of the flavivirus resistance gene.

We also propose to identify possible transcription control signals
on the WNV RNA by investigation of 3' and 5' terminal sequence
conservation among the genomic RNAs of different flaviviruses
and WNV DI RNAs.  The function of these signals will be tested
using a template-dependent in vitro polymerase assay and
genetically engineered RNA transcripts produced from full-length
clones of WNV DI RNA.  Sequence and functional analysis of a
WNV replication-efficient mutant will provide additional
information on the regulatory elements involved in WNV
transcription.

## Key facts

- **NIH application ID:** 3127888
- **Project number:** 5R01AI018382-08
- **Recipient organization:** GEORGIA STATE UNIVERSITY
- **SAM.gov UEI:** MNS7B9CVKDN7
- **Principal Investigator:** Margo A Brinton
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1991
- **Award amount:** $173,103
- **Award type:** 5
- **Project period:** 1983-07-31 → 1993-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3127888

## Citation

> US National Institutes of Health, RePORTER application 3127888, CELLULAR GENE PRODUCT MODULATES FLAVIVIRUS RNA SYNTHESIS (5R01AI018382-08). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3127888. Licensed CC0.

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