Abstract

The object of this research is to understand the role of specific lesions in DNA in effecting cell death. Using antiserum made against UV-irradiated DNA, we have developed a radioimmunoassay capable of detecting photoproducts in the DNA of mammalian cells at biologically relevant dose levels. Depending on the assay conditions, this antiserum can be used to quantify both cyclobutane dimers and Pyr(6-4)Pyo adducts. We now propose to make monoclonal antibodies in order to increase the variety of photoproducts that can be assazed and to analyse intermediary structures formed during DNA repair. The ability of UV-hypersensitive mammalian cell lines to repair various DNA photoproducts will be determined and, using different complementation groups, common repair pathways for different lesions sought. The use of modified UV-DNA/polynucleotides to screen the hybridomas will enable us to isolate potential probes for the analysis of the DNA repair process. The ability to identify intermediate structures will also assist us in elucidating the defect in the mutant cell lines.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
2R01CA024540-07A1
Application #
3166470
Study Section
Chemical Pathology Study Section (CPA)
Project Start
1982-01-01
Project End
1988-08-31
Budget Start
1985-09-30
Budget End
1986-08-31
Support Year
7
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
University of Texas MD Anderson Cancer Center
Department
Type
Organized Research Units
DUNS #
001910777
City
Houston
State
TX
Country
United States
Zip Code
77030

  Publications

Mitchell, D L; Brash, D E; Nairn, R S (1990) Rapid repair kinetics of pyrimidine(6-4)pyrimidone photoproducts in human cells are due to excision rather than conformational change. Nucleic Acids Res 18:963-71
Mitchell, D L; Vaughan, J E; Nairn, R S (1989) Inhibition of transient gene expression in Chinese hamster ovary cells by cyclobutane dimers and (6-4) photoproducts in transfected ultraviolet-irradiated plasmid DNA. Plasmid 21:21-30
Mitchell, D L; Adair, G M; Nairn, R S (1989) Inhibition of transient gene expression in Chinese hamster ovary cells by triplet-sensitized UV-B irradiation of transfected DNA. Photochem Photobiol 50:639-46
Thompson, L H; Mitchell, D L; Regan, J D et al. (1989) CHO mutant UV61 removes (6-4) photoproducts but not cyclobutane dimers. Mutagenesis 4:140-6
Mitchell, D L; Zdzienicka, M Z; van Zeeland, A A et al. (1989) Intermediate (6-4) photoproduct repair in Chinese hamster V79 mutant V-H1 correlates with intermediate levels of DNA incision and repair replication. Mutat Res 226:43-7
Mitchell, D L; Nairn, R S (1989) The biology of the (6-4) photoproduct. Photochem Photobiol 49:805-19
Nairn, R S; Mitchell, D L; Adair, G M et al. (1989) UV mutagenesis, cytotoxicity and split-dose recovery in a human-CHO cell hybrid having intermediate (6-4) photoproduct repair. Mutat Res 217:193-201
Cleaver, J E; Cortes, F; Karentz, D et al. (1988) The relative biological importance of cyclobutane and (6-4) pyrimidine-pyrimidone dimer photoproducts in human cells: evidence from a xeroderma pigmentosum revertant. Photochem Photobiol 48:41-9
Mitchell, D L (1988) The relative cytotoxicity of (6-4) photoproducts and cyclobutane dimers in mammalian cells. Photochem Photobiol 48:51-7
Mitchell, D L (1988) The induction and repair of lesions produced by the photolysis of (6-4) photoproducts in normal and UV-hypersensitive human cells. Mutat Res 194:227-37

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