# ARBOVIRUS REPLICATION IN MOSQUITO AND VERTEBRATE CELLS

> **NIH NIH R01** · UNIV OF MED/DENT NJ-R W JOHNSON MED SCH · 1993 · $229,891

## Abstract

There are almost 400 arthropod-borne viruses, many of which cause serious
human disease, fall into 5 major families. Sindbis virus (SV) in the
family Togaviridae, genus alphavirus, is the subject of this proposal. By
studying the comparative aspects of viral replication in Ae. albopictus
mosquito cells and vertebrate cells, we have been able to isolate mutants
of SV with novel phenotypes: 1) SVLM21, which is able to grow in mosquito
cells which have been deprived of methionine, and have low levels of ado
met, 2) SVmpa, which can grow in mosquito cells which have been treated
with mycophenolic acid, and have low levels of GTP, 3) Svcp which shows
increased cytopathogenicity for mosquito cells, 4) SVcl35 and SVc,158
which are host-restricted in mosquito cells and 5) SVAP15/21 which is
restricted in vertebrate cells. Making use of recombinant DNA technology
and the "infectious" Toto plasmids which contain a CDNA copy of the
Sindbis virus RNA, we are mapping the mutations in these viruses and
determining the nucleotide changes responsible for their phenotypes; this
approach is also making it possible to associate specific biochemical
functions with various of the SV nonstructural (ns) proteins. Thus, most
recently we have been able, using SVLM, CDNA, to associate the viral RNA
methyltransferase with the nonstructural protein, nspl, and by nucleic
acid sequencing to identify two amino acid changes in the SVLM, nspl.
Since we wish eventually to study the structure-function relationships of
nspl we are developing strategies for the construction of expression
vectors which will enable us to produce large amounts of both the
standard and mutant nspl. In a similar fashion we shall use SVmpa to
identify the protein associated with the RNA capping activity (RNA
guanylyltransferase). The significance of these studies lies in the fact
that until now there has been little rigorous evidence which makes it
possible to associate a specific function with any of the four SV ns
proteins. Since SVLM2l and SVMPA, are the only mutants of SV which have
both 1) a change in an ns protein and 2) a phenotype which points to an
alteration in a specific biochemical function, they will be invaluable
for associating at least two functions with specific ns proteins. What we
learn from these studies concerning the SV genome and its replication
strategy may ultimately lead to novel strategies for the control of
medically important viruses, which are transmitted by arthropod vectors.

## Key facts

- **NIH application ID:** 3124272
- **Project number:** 5R01AI005920-31
- **Recipient organization:** UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
- **Principal Investigator:** VICTOR STOLLAR
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1993
- **Award amount:** $229,891
- **Award type:** 5
- **Project period:** 1979-02-01 → 1994-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3124272

## Citation

> US National Institutes of Health, RePORTER application 3124272, ARBOVIRUS REPLICATION IN MOSQUITO AND VERTEBRATE CELLS (5R01AI005920-31). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3124272. Licensed CC0.

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