# COMPOSITION, ASSEMBLY AND REPLICATION OF RNA VIRUSES

> **NIH NIH R01** · UTAH STATE HIGHER EDUCATION SYSTEM--UNIVERSITY OF UTAH · 1988 · $257,110

## Abstract

Our laboratory will study the mechanism(s) involved in the regulation of
the functions of the virus-specific, virion-associated RNA polymerases of
the two minus-stranded RNA animal viruses, vesicular stomatitis virus (VSV)
and influenza A.  Specifically we have designed experiments to examine the
modification(s) of these two enzymes which allows them to switch from
transcription of their genomes into virus-specific mRNAs which are capped,
methylated and polyadenylated and are incomplete copies of the genomic
sequence to a replicative mode which leads to synthesis of full length (+)
and (-) copies of the genome that are assembled during synthesis with
nucleocapsid proteins to form RNP complexes.

We are presently cloning the small subunit of the VSV polymerase (NS) for
expression in E. coli and in animal cells to obtain sufficient amounts of
NS with which to carry out analyses of the phosphorylation sites on NS.  We
will also sequence the cDNA to determine the amino acid sequence of this
protein.  If we can obtain reasonable levels of expression for NS we hope
to use the NS protein for in vitro reconstitution studies with purified
RNPs and L protein.

Using monospecific antisera versus both VSV enzyme subunits (L1, NS1) we
are studying the interaction of this enzyme with its RNP template by
immunoelectron microscopic techniques.

Since we can inhibit influenza transcription (due to its reliance on host
cell mRNA for caps and primers) with actino D, we will establish an in
vitro replication system from cell nuclear extracts for flu by coupling
transcription and translation to drive flu replication in a manner similar
to our in vitro VSV replication system.  Using specific antibodies to the
flu RNA polymerase (PB1, PB2 PA) we will establish whether all three
subunits are involved in flu replication as well as transcription.  In a
similar manner we will study the role, if any, of flu nonstructural protein
(NS1) in vitro replication system in which we can selectively inhibit
transcription (actino D) or replication (cycloheximide) to study the enzyme
modification(s) involved and the role of cellular and/or virus factors in
it.

## Key facts

- **NIH application ID:** 3125161
- **Project number:** 5R01AI012316-13
- **Recipient organization:** UTAH STATE HIGHER EDUCATION SYSTEM--UNIVERSITY OF UTAH
- **SAM.gov UEI:** LL8GLEVH6MG3
- **Principal Investigator:** DONALD F SUMMERS
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1988
- **Award amount:** $257,110
- **Award type:** 5
- **Project period:** 1975-05-01 → 1990-11-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3125161

## Citation

> US National Institutes of Health, RePORTER application 3125161, COMPOSITION, ASSEMBLY AND REPLICATION OF RNA VIRUSES (5R01AI012316-13). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3125161. Licensed CC0.

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