This is a revised application which is focused on the analysis of the mechanism of dRpase activity in bacteria, and the identification of corresponding activities in yeast.
The specific aims are: (1) A novel plasmid assay construct will be used to analyze the role of the several known dRpase activities in bacterial DNA base excision repair. The plasmid assay construct will also be used to determine if the repair process involves replacement of one or several nucleotides. (2) Interactions between several proteins that function as part of the DNA base excision repair pathway will be investigated by use of the yeast two- hybrid system, and where appropriate by oligo-histidine affinity chromatography. Interactions will also be studied with purified repair enzymes and the plasmic assay system of SA1, in vitro, by analysis of repair rates as a function of added protein co-factors. (3) Characterization and purification of a eukaryotic dRpase activity in the yeast S. pombe. The yeast enzyme will be purified and subsequently subjected to peptide sequencing so as to yield oligonucleotide probes which can be used to screen a yeast cDNA library for the corresponding mRNA. It is proposed that this would allow for the sequencing, cloning, and possible source to identify a homologous human allele.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA052025-08
Application #
2700438
Study Section
Chemical Pathology Study Section (CPA)
Program Officer
Pelroy, Richard
Project Start
1990-04-01
Project End
2001-04-30
Budget Start
1998-05-01
Budget End
1999-04-30
Support Year
8
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Albert Einstein College of Medicine
Department
Radiation-Diagnostic/Oncology
Type
Schools of Medicine
DUNS #
009095365
City
Bronx
State
NY
Country
United States
Zip Code
10461
Lombardo, Mary-Jane; Aponyi, Ildiko; Ray, Mellanie P et al. (2003) xni-deficient Escherichia coli are proficient for recombination and multiple pathways of repair. DNA Repair (Amst) 2:1175-83
Sandigursky, M; Franklin, W A (1998) Exonuclease IX of Escherichia coli removes 3' phosphoglycolate end groups from DNA. Radiat Res 150:609-11
Li, W X; Franklin, W A (1998) Radiation- and heat-induced apoptosis in PC-3 prostate cancer cells. Radiat Res 150:190-4