# DNA DAMAGE IN ONCOGENE ACTIVATION

> **NIH NIH R01** · OHIO STATE UNIVERSITY · 1989 · $137,688

## Abstract

The aim of this proposal is to determine if specific DNA base
alterations induced by environmental carcinogens can cause
critical mutations in a target gene leading to its activation and
subsequent initiation of neoplastic process.  The objectives are:
(i) to understand the role of mutagenic DNA base modifications
specifically incorporated within a defined region of ras proto-
oncogene without subjecting the target cells to direct carcinogen
exposure; (ii) identify the sequence changes of the modified region
responsible for altered gene function and cellular phenotype; and
(iii) compare the response of NIH/3T3 and human cells in in vitro
transformation by transfection with specifically altered cloned
genes.  These studies will provide an insight on the role of specific
adduct and site related critical mutational activation of normal
cellular genes in the process of neoplastic cellular transformation
and lead to the development of a more relevant assay system
using human cells as the recepeints of transfected oncogenes.

The approach to be employed in this study is as follows.  The
cloned ras gene (plasmid pbc-N1) will be site specifically modified
to solely contain principal mutagenic lesions of DNA alkylation
(O6-ethylguanine or O4-ethylthymine) targeted within a specific
coding regions of the gene by AMV DNA polymerase.  The effect
of each of the lesions will be studied in the induction of mutations
at critical sites of ras gene and their relative potency in
conferring the transforming properties upon this gene.  Mutated
plasmid will be introduced into the mouse NIH/3T3 and human
fibroblast cells by calcium phosphate mediated DNA transfection
and co-transfection with other genes.  Neoplastic phenotype
conferred by establishment of modified gene in the transformants
will be determined by scoring the morphologically altered foci,
anchorage-independent cellular growth and tumorigenicity in nude
mice.

## Key facts

- **NIH application ID:** 3178312
- **Project number:** 5R01CA039397-03
- **Recipient organization:** OHIO STATE UNIVERSITY
- **SAM.gov UEI:** DLWBSLWAJWR1
- **Principal Investigator:** ALTAF A WANI
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1989
- **Award amount:** $137,688
- **Award type:** 5
- **Project period:** 1987-09-30 → 1991-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3178312

## Citation

> US National Institutes of Health, RePORTER application 3178312, DNA DAMAGE IN ONCOGENE ACTIVATION (5R01CA039397-03). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3178312. Licensed CC0.

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