# HISTONE DEPOSITION IN VIRALLY-INFECTED AND TRANSFORMED

> **NIH NIH R01** · MEDICAL COLLEGE OF WISCONSIN · 1988 · $89,759

## Abstract

We plan to continue our studies into the mechanism of histone
deposition during DNA replication in SV40 infected and
transformed cells.  We will further characterize the sites in the
genome where the deposition of newly synthesized H2A, H2B
occurs.  We will utilize newly developed procedures of
immunoprecipitation of ubiquinated histone complexes
(crosslinked octamers) to determine whether the exchange
phenomenon relates to the potential instability of a ubiquitin
moiety within  a nucleosome.  We will use newly developed
procedures of nick translation of nuclei with biotinylated
nucleotide to isolate active genes on avidin-agarose.  We will
continue our studies into the exchange phenomenon using density
labeled amino acids (15N, 13C, 2H) to label histones, crosslink the
histones within the nucleosome into an octameric complex with
formaldehyde and analyze the density of the complex.  Because of
the reversibility of this crosslink, the individual histones can be
charaterized within the density fractionated octamers.  We will
try to perturb the exchange process by varying tempature (cells
that grow between 4 and 37 degrees Centigrade) and in treatment
with metabolic inhibitors such as actinomycin D.  We will
continue our studies into the role of superhelical density in
regulating histone deposition.  Histone deposition on new DNA is
extensively inhibited in cells treated with the intercalating agent,
chloroquin.  This is shown by crosslinking whole cells with
formaldehyde, isolating the chromatin and fractionating on
density gradients.  We will determine the superhelical density and
continuity of this new DNA.  We will test whether a potential
lethal effect of anticancer drugs (those involved in protein
mediated strand breakage) is through the inhibition of histone
deposition on newly replicated DNA.  We will utilize in vitro
deposition procedures with polyglutamic acid:histone and DNA
(supercoiled and relaxed) to measure rates of histone deposition
(newly synthesized and old) and to measure the effect of
nucleosome structure on the interaction of transcription factor,
ICP4, with targe DNA sequences.  These combined studies will
provide a clearer understanding into the role of histones in the
regulation of replication and transcription.

## Key facts

- **NIH application ID:** 3173383
- **Project number:** 5R01CA035829-05
- **Recipient organization:** MEDICAL COLLEGE OF WISCONSIN
- **SAM.gov UEI:** E8VWJXMMUQ67
- **Principal Investigator:** VAUGHN E JACKSON
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1988
- **Award amount:** $89,759
- **Award type:** 5
- **Project period:** 1983-07-01 → 1990-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3173383

## Citation

> US National Institutes of Health, RePORTER application 3173383, HISTONE DEPOSITION IN VIRALLY-INFECTED AND TRANSFORMED (5R01CA035829-05). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3173383. Licensed CC0.

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