# MOLECULAR AND GENETIC ANALYSIS OF PARVOVIRUS REPLICATION

> **NIH NIH R01** · YALE UNIVERSITY · 1987 · $241,811

## Abstract

Minute virus of mice (MVM) is an autonomous parvovirus which exhibits
teratogenic and tumor-suppressive activity in vivo.  We propose to define
precisely the mechanism by which MVM replicates its genome and to elucidate
the functions that both viral and cellular proteins perform in the
replication process.  The primary gene products of MVM will be enumerated,
and the specific transcripts coding for them identified, by in vitro
translation of infected cell RNA purified by hybridization to cloned
segments of the viral genome.  The coding regions which are spliced
together to produce these polypeptides will be determined by cloning and
sequencing cDNA copies of each vira mRNA species.  The unique and shared
regions of each viral gene will then be located within the entire DNA
sequence of MVM, which has recently been determined.  Specific mutations
will be introduced into unique coding sequences in an infectious viral
genome which as been cloned into a bacterial plasmid.  Similarly, mutations
will be introduced into the terminal regions of the genome, known to
contain all of the cis-acting functions essential for viral DNA replication
and progeny virion maturation.  Mutant plasmids will then be transfected
into appropriate host cells and the biological functions of these regions
in the viral life cycle analyzed.  A parallel approach, designed also to
identify cellular factors essential for viral DNA synthesis, will examine
the components of viral DNA replication complexes.  These will be isolated
by both conventional purification schemes and by affinity chromatography on
avidin-Sepharose following in vivo pulse-labelling with biotinyl
nucleotides.  The proteins present in these complexes will be separated and
characterized as to genetic origin using immunological reagents, 2-D gel
electrophoresis and peptide-mapping.  In addition, each component of the
complex will be evaluated for specific enzymatic activities.  Finally,
monoclonal antibodies against each will be prepared and used to purify the
protein and further investigate its function.  These purified components
will be used to reconstruct the viral DNA replication machinery in vitro.
Since parvoviruses have a limited genetic capacity and rely extensively
upon host cell functions for their growth, we anticipate that our studies
will also provide some basic insights into the mechanism of eukaryotic cell
DNA replication.

## Key facts

- **NIH application ID:** 3129438
- **Project number:** 5R01AI019973-05
- **Recipient organization:** YALE UNIVERSITY
- **SAM.gov UEI:** FL6GV84CKN57
- **Principal Investigator:** DAVID C WARD
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1987
- **Award amount:** $241,811
- **Award type:** 5
- **Project period:** 1983-04-01 → 1988-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3129438

## Citation

> US National Institutes of Health, RePORTER application 3129438, MOLECULAR AND GENETIC ANALYSIS OF PARVOVIRUS REPLICATION (5R01AI019973-05). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3129438. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*