Abstract

Cells derived from patients with xeroderma pigmentosum, who are predisposed to sunlight induced skin cancer, are unable to catalyze incision of DNA damaged by light as well as by a variety of structurally unrelated carcinogens. The three structural genes responsible for this enzymes reaction in E. coli have been cloned and their gene product uvrA, uvrB and uvrC proteins have been isolated determinations and for their use as antigens. These three genes are to be separately introduced into the heterologous composite pBR322-SV40 plasmid, pSV2-gpt, containing the E. coli gene (gpt) which encodes the xanthine guanine phosphoribosyl transferase (XGPRT) gene (Mulligan, R.C. and Berg, P., (1980) Science 209, 1427). Cotransfection of the uvr genes with gpt into monkey kidney cells (BSC-1) and human skin fibroblasts in a medium selectively supporting growth of only those cells expressing gpt greatly facilitates selection for those cells cotransfected with uvr genes. Transfected cells will be screened for XGPRTase, HPRTase, gpt mRNA, uvr mRNA uvr DNA fluorescene. The reversion of those repair deficient cells expressing uvr genes to a state of repair proficiency will be determined.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM031110-04
Application #
3279049
Study Section
Biochemistry Study Section (BIO)
Project Start
1982-07-01
Project End
1987-06-30
Budget Start
1985-07-01
Budget End
1986-06-30
Support Year
4
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
Johns Hopkins University
Department
Type
Schools of Public Health
DUNS #
045911138
City
Baltimore
State
MD
Country
United States
Zip Code
21218

  Publications

Wei, Q; Matanoski, G M; Farmer, E R et al. (1995) DNA repair capacity for ultraviolet light-induced damage is reduced in peripheral lymphocytes from patients with basal cell carcinoma. J Invest Dermatol 104:933-6
Wei, Q; Matanoski, G M; Farmer, E R et al. (1994) DNA repair related to multiple skin cancers and drug use. Cancer Res 54:437-40
Wang, J; Mueller, K L; Grossman, L (1994) A mutational study of the C-terminal zinc-finger motif of the Escherichia coli UvrA protein. J Biol Chem 269:10771-5
Wei, Q; Matanoski, G M; Farmer, E R et al. (1994) DNA repair and susceptibility to basal cell carcinoma: a case-control study. Am J Epidemiol 140:598-607
Wei, Q; Matanoski, G M; Farmer, E R et al. (1994) Vitamin supplementation and reduced risk of basal cell carcinoma. J Clin Epidemiol 47:829-36
Wang, J; Grossman, L (1993) Mutations in the helix-turn-helix motif of the Escherichia coli UvrA protein eliminate its specificity for UV-damaged DNA. J Biol Chem 268:5323-31
Wei, Q; Matanoski, G M; Farmer, E R et al. (1993) DNA repair and aging in basal cell carcinoma: a molecular epidemiology study. Proc Natl Acad Sci U S A 90:1614-8
Thiagalingam, S; Grossman, L (1993) The multiple roles for ATP in the Escherichia coli UvrABC endonuclease-catalyzed incision reaction. J Biol Chem 268:18382-9
Athas, W F; Hedayati, M A; Matanoski, G M et al. (1991) Development and field-test validation of an assay for DNA repair in circulating human lymphocytes. Cancer Res 51:5786-93
Claassen, L A; Grossman, L (1991) Deletion mutagenesis of the Escherichia coli UvrA protein localizes domains for DNA binding, damage recognition, and protein-protein interactions. J Biol Chem 266:11388-94

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