These studies will examine the molecular mechanisms for repair of radiation-induced DNA damage in human tissue culture cells. The project focuses on determining whether ionizing and ultraviolet radiation may induce gene expression. Although DNA damage inducible functions have been identified and extensively studied in E. coli (the SOS system), the evidence for inducible functions in mammalian cells is incomplete and largely based on induction, mutagenesis and host cell reactivation of viral probes. We propose to detect and clone genes inducible by ionizing or UV radiation by using two sensitive and direct techniques: 1) detection of radiation induced alterations in specific mRNA contnent using hybridization of cDNAs against a genomic library, and 2) induction of selectable markers fused into eukaryotic transcriptional units via transfection. Identification and cloning of inducible genes will lead to further elucidation of the pathways for repair of radiation-induced DNA damage in eukaryotes. This may have broad implications in mutagenesis and carcinogenesis.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Research Project (R01)
Project #
5R01ES003446-02
Application #
3250735
Study Section
Chemical Pathology Study Section (CPA)
Project Start
1984-07-01
Project End
1987-06-30
Budget Start
1985-07-01
Budget End
1986-06-30
Support Year
2
Fiscal Year
1985
Total Cost
Indirect Cost
Name
U.S. Uniformed Services University of Health Science
Department
Type
Schools of Medicine
DUNS #
City
Bethesda
State
MD
Country
United States
Zip Code
20814
Radany, E H; Nguyen, H T; Minton, K W (1987) Activities involved in base excision repair of bacteriophage T4 and lambda DNA in vivo. Mol Gen Genet 209:83-9
Love, J D; Nguyen, H T; Or, A et al. (1986) UV-induced interstrand cross-linking of d(GT)n.d(CA)n is facilitated by a structural transition. J Biol Chem 261:10051-7
Attri, A K; Minton, K W (1986) An automated method for determining buoyant density of nucleic acids using a preparative ultracentrifuge. Anal Biochem 159:88-95
Vivino, A A; Smith, M D; Minton, K W (1986) A DNA damage-responsive Drosophila melanogaster gene is also induced by heat shock. Mol Cell Biol 6:4767-9
Love, J D; Minton, K W (1985) Screening of lambda library for differentially expressed genes using in vitro transcripts. Anal Biochem 150:429-41
Minton, K W (1985) The triple helix: a potential mechanism for gene regulation. J Exp Pathol 2:135-48