The major thrust of this application is to understand the molecular details of how various cancer therapeutic modalities affect DNA. We have developed a simplified general approach to the study of the physicochemical and biochemical consequences of structural lesions in DNA, using DNA oligomers containing a specific structural lesion at a single site at 100% frequency. We will use this general approach for the study of several important DNA lesions including: (1) the lesion introduced through misincorporation of DHPG, an exciting new anti-viral agent, (2) the hydroxymethal damage to DNA, and (3) the 5-azacytidine lesion introduced by misincorporation. We will study biochemical effects on processes related to DNA replication such as chain elongation, ligation, and movement of a complete replication fork past a lesion site. Using NMR spectroscopy, we will investigate the physicochemical consequences of these lesions. Through construction of computer graphic models we will investigate the detailed structural consequences of lesions in DNA.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
2R01CA042300-03
Application #
3183388
Study Section
Experimental Therapeutics Subcommittee 2 (ET)
Project Start
1985-07-01
Project End
1992-11-30
Budget Start
1987-12-01
Budget End
1988-11-30
Support Year
3
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Yale University
Department
Type
Schools of Medicine
DUNS #
082359691
City
New Haven
State
CT
Country
United States
Zip Code
06520
Marshalko, S J; Schweitzer, B I; Beardsley, G P (1995) Chiral chemical synthesis of DNA containing (S)-9-(1,3-dihydroxy-2-propoxymethyl)guanine (DHPG) and effects on thermal stability, duplex structure, and thermodynamics of duplex formation. Biochemistry 34:9235-48
Mikita, T; Beardsley, G P (1994) Effects of arabinosylcytosine-substituted DNA on DNA/RNA hybrid stability and transcription by T7 RNA polymerase. Biochemistry 33:9195-208
Schweitzer, B I; Mikita, T; Kellogg, G W et al. (1994) Solution structure of a DNA dodecamer containing the anti-neoplastic agent arabinosylcytosine: combined use of NMR, restrained molecular dynamics, and full relaxation matrix refinement. Biochemistry 33:11460-75
Sokoloski, J A; Beardsley, G P; Sartorelli, A C (1989) Induction of HL-60 leukemia cell differentiation by the novel antifolate 5,10-dideazatetrahydrofolic acid. Cancer Res 49:4824-8
Mikita, T; Beardsley, G P (1988) Functional consequences of the arabinosylcytosine structural lesion in DNA. Biochemistry 27:4698-705
Spigelman, Z; Duff, R; Beardsley, G P et al. (1988) 2',3'-Dideoxyadenosine is selectively toxic for TdT-positive cells. Blood 71:1601-8
Beardsley, G P; Mikita, T; Klaus, M M et al. (1988) Chemical synthesis of DNA oligomers containing cytosine arabinoside. Nucleic Acids Res 16:9165-76
Kremer, A B; Mikita, T; Beardsley, G P (1987) Chemical consequences of incorporation of 5-fluorouracil into DNA as studied by NMR. Biochemistry 26:391-7
Clark, J M; Joyce, C M; Beardsley, G P (1987) Novel blunt-end addition reactions catalyzed by DNA polymerase I of Escherichia coli. J Mol Biol 198:123-7