Abstract

These studies will examine some of the enzymatic properties of endonucleas III and IV of E. coli and endonuclease V of bacteriophage T4. Purification of these enzymes to homogeneity will allow us to define exact substrates and products for the reactions catalyzed by endonuclease III on DNA containing modified pyrimidine residues, by endonuclease IV on apurinic and apyrimidinic DNA, and by endonucleases III, IV and V on DNA containing deletion loops. In vitro site directed mutagenesis will be used to obtain mutants for endonuclease III. Purification and characterization of the mutant endonuclease III enzymes will help define the number of active sites within the protein and DNA sequencing of wild-type and mutant genes may allow us to define the active sites. Base excision repair may be one of the major pathways by which cells repair damaged bases in DNA. Enzymatic activities for such repair have been found in a large number of prokaryotes and eukaryotes. A careful study of some of these enzymes will be useful in defining the exact fashion in which cells repair DNA damaged by ultraviolet and X-irradiation and chemicals. Such treatments are used to treat cancer, and are also known to promote cancer.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM033346-04
Application #
3282948
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1984-12-01
Project End
1989-11-30
Budget Start
1987-12-01
Budget End
1989-11-30
Support Year
4
Fiscal Year
1988
Total Cost
Indirect Cost

  Institution

Name
State University of New York at Albany
Department
Type
Schools of Arts and Sciences
DUNS #
City
Albany
State
NY
Country
United States
Zip Code
12222

  Publications

Mazumder, A; Gerlt, J A; Absalon, M J et al. (1991) Stereochemical studies of the beta-elimination reactions at aldehydic abasic sites in DNA: endonuclease III from Escherichia coli, sodium hydroxide, and Lys-Trp-Lys. Biochemistry 30:1119-26
Boorstein, R J; Hilbert, T P; Cunningham, R P et al. (1990) Formation and stability of repairable pyrimidine photohydrates in DNA. Biochemistry 29:10455-60
Doetsch, P W; Cunningham, R P (1990) The enzymology of apurinic/apyrimidinic endonucleases. Mutat Res 236:173-201
Mueller, J E; Newton, C J; Jensch, F et al. (1990) Resolution of Holliday junction analogs by T4 endonuclease VII can be directed by substrate structure. J Biol Chem 265:13918-24
Boorstein, R J; Hilbert, T P; Cadet, J et al. (1989) UV-induced pyrimidine hydrates in DNA are repaired by bacterial and mammalian DNA glycosylase activities. Biochemistry 28:6164-70
Cunningham, R P; Asahara, H; Bank, J F et al. (1989) Endonuclease III is an iron-sulfur protein. Biochemistry 28:4450-5
Asahara, H; Wistort, P M; Bank, J F et al. (1989) Purification and characterization of Escherichia coli endonuclease III from the cloned nth gene. Biochemistry 28:4444-9
Mueller, J E; Kemper, B; Cunningham, R P et al. (1988) T4 endonuclease VII cleaves the crossover strands of Holliday junction analogs. Proc Natl Acad Sci U S A 85:9441-5
Gossett, J; Lee, K; Cunningham, R P et al. (1988) Yeast redoxyendonuclease, a DNA repair enzyme similar to Escherichia coli endonuclease III. Biochemistry 27:2629-34
Saporito, S M; Cunningham, R P (1988) Nucleotide sequence of the nfo gene of Escherichia coli K-12. J Bacteriol 170:5141-5

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