# MOLECULAR GENETICS OF HSV DNA POLYMERASE GENE

> **NIH NIH R01** · HARVARD MEDICAL SCHOOL · 1993 · $252,334

## Abstract

We wish to gain a comprehensive understanding of the regulation and
function of the herpes simplex virus (HSV) DNA polymerase (pol) gene.
HSV Pol is both an excellent model for eukaryotic replicative polymerases
and an important target for antiviral drugs.  HSV and other herpesviruses
are important pathogens, especially in immunocompromised individuals such
as AIDS patients and transplant and chemotherapy recipients.  Drug
resistance, due in part to pol mutations, is a growing problem.
     The mechanisms regulating the translational shut-off of pol gene
expression and its importance to the virus will be analyzed through
mutations that fail to down-regulate translation of pol mRNA.  Studies of
Pol function are facilitated by the availability of quantities of
purified Pol from recombinant baculovirus-infected insect cells.  The
enzyme will be proteolytically cleaved into domains, which will be tested
for various Pol activities.  The structure of Pol and its domains will be
examined by X-ray crystallography.  Pol interactions with nucleic acids
will be studied with spectroscopic, mobility shift and footprinting
methods.  Amino acids that bind deoxynucleoside substrates will be
identified by affinity labeling.  Genetic correlates to protein domains
will be developed by analysis of mutations that exhibit intragenic
complementation.  The roles of conserved regions of Pol that are altered
by acyclovir-resistance mutations will be investigated to determine which
steps in the mechanism of acyclovir-triphosphate inhibition of Pol each
mutation blocks.  Amino acids hypothesized to be within active sites for
the 3'-5' exonuclease and RNase H/5'3' exonuclease will be altered by
site-specific mutagenesis and the importance of these residues for
various Pol enzyme activities and for virus growth will be tested.  A
signal required for the localization of Pol to prereplicative sites in
HSV-infected cells, which is dependent upon the viral single-stranded DNA
binding protein, ICP8, will be sought by studies of mutations that
prevent localization.  The importance to the virus of this signal and of
a domain that binds to the polymerase accessory protein, UL42, will be
assessed by construction of HSV mutants.  These studies should permit a
detailed picture of the regulation and functional domains of Pol both in
vitro and in infected cells and may uncover novel features of polymerases
and new strategies for antiviral drug development.

## Key facts

- **NIH application ID:** 3129277
- **Project number:** 5R01AI019838-11
- **Recipient organization:** HARVARD MEDICAL SCHOOL
- **SAM.gov UEI:** JDLVAVGYJQ21
- **Principal Investigator:** DONALD M COEN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1993
- **Award amount:** $252,334
- **Award type:** 5
- **Project period:** 1983-04-01 → 1996-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3129277

## Citation

> US National Institutes of Health, RePORTER application 3129277, MOLECULAR GENETICS OF HSV DNA POLYMERASE GENE (5R01AI019838-11). Retrieved via MORVS 2026-10-24 from https://api.morvs.ai/grant/nih/3129277. Licensed CC0.

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