# SEQUENCE-SPECIFIC INCORPORATION OF ANALOGS INTO SV40 DNA

> **NIH NIH R01** · UNIVERSITY OF MICHIGAN AT ANN ARBOR · 1987 · $73,061

## Abstract

Many anticancer and antiviral agents are analogs of purines or pyrimidines
that are thought to act by being incorporated into DNA.  Most of the
studies performed to date concerning the mechanism by which these drugs act
have used DNA into which the analog has been randomly incorporated.  For
one such analog, 6-thioguanine (TG), we have performed experiments that
suggest that the action of this drug may depend on its being incorporated
into a specific DNA sequence.  The overall objective of the proposed work
is to determine if such "sequence-specific incorporation" is necessary for
the action of Tg, and also of three other analogs which are believed to act
via incorporation into DNA.  This objective will be met by achieving the
following specific goals:  1.  Construction of specifically substituted
viral DNA molecules.  The eukaryotic virus SV40 will be the model system
for these studies.  SV40 DNA contains a sequence at its origin of
replication whose integrity has been shown to be critical for initiation of
viral DNA synthesis.  Analog substitutions will be performed either
collectively within this sequence or in other regions of the viral genome
known to be insensitive to mutation.  In addition to TG, substitutions will
be performed using ara-C, ara-A or bromodeoxyuridine.  2.  Testing the
biological effects of sequence-specific substitution.  Once the contructed
molecules are in hand their ability to function will be tested in two
systems.  First, infectivity of these species will be tested by
DEAE-dextran mediated transfection.  Second, the ability of the substituted
molecules to function as templates in a recently reported cell-free system
will be assayed.  3.  Testing the protein binding properties of substituted
DNA.  The virally-coded "T-antigen" (T-ag), whose function is critical for
initiation of replication, binds to the site within the origin where
specific substitutions will be made.  The affinity of analog-substituted
monlecules for T-ag will be measured by means of an immunoprecipitation
procedure using an anti-T-ag monoclonal antibody.

## Key facts

- **NIH application ID:** 3179991
- **Project number:** 5R01CA040260-03
- **Recipient organization:** UNIVERSITY OF MICHIGAN AT ANN ARBOR
- **SAM.gov UEI:** GNJ7BBP73WE9
- **Principal Investigator:** Jonathan Maybaum
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1987
- **Award amount:** $73,061
- **Award type:** 5
- **Project period:** 1985-08-01 → 1988-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3179991

## Citation

> US National Institutes of Health, RePORTER application 3179991, SEQUENCE-SPECIFIC INCORPORATION OF ANALOGS INTO SV40 DNA (5R01CA040260-03). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3179991. Licensed CC0.

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