# CYTOMEGALOVIRUS DNA REPLICATION AND INVERSION

> **NIH NIH R01** · STANFORD UNIVERSITY · 1988 · $168,028

## Abstract

This proposal will study molecular events and gene products
involved in genome replication, circularization, inversion and
cleavage/packaging of herpesviruses with emphasis on a beta
herpesvirus, human cytomegalovirus (CMV).  Viral and host cell
proteins that bind to or enzymatically act on the CMV a sequence,
which is the cis-acting signal involved in genome circularization
inversion and cleavage/packaging, will be isolated and their
recognition sites will be determined.  The genes for these viral
proteins will be mapped on the viral genome.  As a
complementary approach to identify viral gene products involved
in replication functions, the genes encoding viral nonstructural
nuclear proteins will be mapped on the viral genome.  Monoclonal
and polyclonal antibodies specific for nuclear proteins will be
generated in mice and viral genes will be identified via a lambda
gtll library expressing random CMV open reading frames.
Functional requirement for identified genes will be determined
using insertional mutagenesis of the CMV genome.  CMV insertion
mutants will be constructed using E. coli beta galactosidase as a
genetic marker.  Gene products provided in trans from engineered
retrovirus vectors to complement insertion mutations in essential
CMV genes.  These experiments will identify viral proteins that
have specific functions in replication or have a nuclear
localization and are nonstructural consistent with replication
function.  The genes encoding these proteins will be mapped on
the viral genome and insertional-mutants will be derived.  The
mutants will enable an unambiguous determination of the
functional significance of these gene products in replication.

One area of herpesvirus replication will be studied using herpes
simplex virus:  the role of the HSV-1 DNA replication origin
specific binding protein (ori bp) as a trans-acting function in DNA
replication initiation.  The ori bp gene will be mapped and
conditionally lethal mutants will be derived to demonstrate the
role of this gene product in HSV-1 DNA replication.  The
functional significance of the binding site for ori bp within oriS of
HSV-1 will be assessed by site directed mutagenesis.  OriS with
one, two or zero ori bp sites will be compared for replication
efficiency in competition assays.  The role of the multiple DNA
replication origins on the viral genome will be directly
investigated by introducing modified versions oriS in place of
existing origins.

This research effort will provide a molecular understanding of the
process and control of herpesvirus replication functions.  This
knowledge will impact directly, in the medical terms through the
identification of targets for antiviral development.  The
understanding gained through the proposed research will enable
characterization of more subtle viral functions, for example those
involved in latency in future years.

## Key facts

- **NIH application ID:** 3129722
- **Project number:** 5R01AI020211-05
- **Recipient organization:** STANFORD UNIVERSITY
- **SAM.gov UEI:** HJD6G4D6TJY5
- **Principal Investigator:** EDWARD S. Edward S Mocarski
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1988
- **Award amount:** $168,028
- **Award type:** 5
- **Project period:** 1984-03-01 → 1992-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3129722

## Citation

> US National Institutes of Health, RePORTER application 3129722, CYTOMEGALOVIRUS DNA REPLICATION AND INVERSION (5R01AI020211-05). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3129722. Licensed CC0.

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