# REGULATION OF GENE EXPRESSION IN HSV INFECTED CELLS

> **NIH NIH R01** · MEDICAL COLLEGE OF WISCONSIN · 1988 · $141,850

## Abstract

The viral polypeptide designated ICP4 is synthesized during the
immediate early phase of productive infection by herpes simples
virus (HSV).  ICP4 is a positive regulator of early gene
transcription and a negative regulator of immediate early gene
transcription.  Native ICP4 has been partially purified and shown
to exist as a dimer which associates directly or indirectly with
specific sequences in double-stranded DNA.  The specific aims of
this investigation are (i) to obtain a homogeneous preparation of
ICP4; (ii) to determine what factors are required to stabilize the
association between ICP4 and DNA; (iii) to identify the location
and sequence of nucleotides recognized by ICP4 (or associated
factors) in the viral genome; (iv) to characterize potential
catalytic activities associated with ICP4; (v) to map the location
of functional domains in ICP4; (vi) to establish the mechanism for
ICP4-mediated repression of transcription; and (vii) to investigate
the mechanism for ICP4-mediated induction of transcription.

Purification of ICP4 will be achieved by DNA affinity and
immunoaffinity chromatography.  Both radioimmunoassays and
DNA binding assays will be used to quantitate the recovery of
ICP4 during purification.  The stability of ICP4:DNA complexes
will be analyzed by filter binding assays.  A library of cloned
fragments of HSV DNA will be screened for the presence of ICP4
binding sites by DNA immunoassays.  The nucleotide sequences in
the binding sites will be identified by DNase footprinting.  A
computer-assisted homology search will be used to locate related
sequences in viral and cellular DNAs.  The role that these ICP4
binding sites play in the control of gene transcription will be
examined by in vitro transcription assays using templates which
contain modified forms of an ICP4 binding site.  Fragments of the
ICP4 polypeptide will be produced in E. coli by insertion of
subgenic portions of the ICP4 gene into bacterial expression
vectors.  These polypeptide fragments will be reacted with
monoclonal antibodies against ICP4 and with radiolabeled DNA in
binding reactions designed to map the location of active domains
in ICP4.  These studies will focus on the long range goal of this
investigation, which is to understand the mechanisms which
regulate gene expression in mammalian cells.  Because regulation
of gene expression is a fundamental process that controls the
growth, development, and differentiation of human cells, it is
important that these mechanisms be understood.

## Key facts

- **NIH application ID:** 3127050
- **Project number:** 5R01AI017246-08
- **Recipient organization:** MEDICAL COLLEGE OF WISCONSIN
- **SAM.gov UEI:** E8VWJXMMUQ67
- **Principal Investigator:** KENT W WILCOX
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1988
- **Award amount:** $141,850
- **Award type:** 5
- **Project period:** 1980-07-01 → 1990-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3127050

## Citation

> US National Institutes of Health, RePORTER application 3127050, REGULATION OF GENE EXPRESSION IN HSV INFECTED CELLS (5R01AI017246-08). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3127050. Licensed CC0.

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