# IN VITRO FUNCTION OF DNA CONTAINING CARCINOGEN ADDUCTS

> **NIH NIH R01** · WAYNE STATE UNIVERSITY · 1988 · $75,044

## Abstract

The objective is to continue to investigate the detailed effects of the
bulky carcinogenic adducts, 2-aminofluorene (AF), N-acetyl-2-aminofluorene
(AAF), and benzo(a)pyrene (BP), on DNA replication.  Our approach has been
to study the effect of these adducts on not just a DNA polymerase alone,
but, instead, to determine how a purified in vitro replication system
responds to this type of damage.  We propose to determine how the ancillary
proteins, required for DNA replication in vivo, affect synthesis using
single- or double-stranded DNA templates containing carcinogenic DNA
adducts and to characterize the effect of such adducts on the fidelity of
DNA synthesis, DNA unwinding, RNA transcription, DNA initiation, and
protein-nucleic acid interactions.  The mechanism by which E. coli single
stranded DNA binding protein (SSB) stimulates DNA synthesis by the T7 DNA
polymerase on native and carcinogen-modified DNA will be determined and
this effect will be compared with the results obtained with other DNA
polymerases.  These experiments will utilize not only randomly modified
single- and double-stranded DNA templates, but also DNA containing a
site-specific AF and AAF adducts, which we have recently prepared.  Using
these site-specifically modified templates, the frequency of
misincorporation during trans-lesion synthesis by DNA polymerases in the
presence and absence of SSB or other single-stranded DNA binding proteins
will be determined.  DNA synthesis by the T7 DNA polymerase and gene 4
protein on double-stranded DNA containing AF, AAF, and BP adducts will be
characterized and these studies will be subsequently extended to
site-specifically modified templates.  DNA molecules containing a T7 class
III RNA polymerase promoter and containing an AF or AAF adduct located a
unique site will be prepared and used to further characterize the effect of
DNA adducts on RNA synthesis and on the initiation of DNA replication by
the T7 RNA polymerase, T7 DNA polymerase, and gene 4 protein.  Finally, we
will determine the effect of DNA adducts on the interactions between DNA
and single-stranded DNA binding proteins, T7 RNA polymerase, or the Uvr ABC
endonuclease.

## Key facts

- **NIH application ID:** 3173012
- **Project number:** 5R01CA035451-05
- **Recipient organization:** WAYNE STATE UNIVERSITY
- **SAM.gov UEI:** M6K6NTJ2MNE5
- **Principal Investigator:** Louis J Romano
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1988
- **Award amount:** $75,044
- **Award type:** 5
- **Project period:** 1983-08-01 → 1990-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3173012

## Citation

> US National Institutes of Health, RePORTER application 3173012, IN VITRO FUNCTION OF DNA CONTAINING CARCINOGEN ADDUCTS (5R01CA035451-05). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3173012. Licensed CC0.

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