This Program combines two powerful analytical approaches to discover the causes of spontaneous mutations in human cells, yeasts and bacteria. The first approach involves a major technical advance in mutational spectrometry which permits measurement of predominate point mutations without reference to changes in phenotype. The second approach is the use of biochemical genetics aimed with the many known mutant genes for DNA replication and DNA repair in yeasts and bacteria and the growing set of such genes known in humans. By using such mutants and observing changes in spontaneous mutational spectra we expect to gain important new mechanistic understanding. A series of specific hypotheses regarding the role of DNA manipulation or endogenous DNA damage are presented throughout five complementary projects. The Program's direction is strongly influenced by a growing body of evidence that mutations in human germinal and somatic cells contain a significant fraction of mutations which appear to be spontaneous in origin. The Program's five projects share: -a common focus on spontaneous mutation mechanisms, -a set of common DNA sequences found in the human genome and genetically engineered into yeasts and bacteria -a common technological tool-constant denaturant gradient electrophoresis/high fidelity PCR -applied to obtain point mutational spectra.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Research Program Projects (P01)
Project #
2P01ES003926-11
Application #
2153474
Study Section
Special Emphasis Panel (SRC)
Project Start
1985-09-27
Project End
1998-08-31
Budget Start
1995-09-30
Budget End
1996-08-31
Support Year
11
Fiscal Year
1995
Total Cost
Indirect Cost
Name
Massachusetts Institute of Technology
Department
Miscellaneous
Type
Other Domestic Higher Education
DUNS #
City
Cambridge
State
MA
Country
United States
Zip Code
02139
Memisoglu, A; Samson, L (2000) Contribution of base excision repair, nucleotide excision repair, and DNA recombination to alkylation resistance of the fission yeast Schizosaccharomyces pombe. J Bacteriol 182:2104-12
Wyatt, M D; Samson, L D (2000) Influence of DNA structure on hypoxanthine and 1,N(6)-ethenoadenine removal by murine 3-methyladenine DNA glycosylase. Carcinogenesis 21:901-8
Opperman, T; Murli, S; Smith, B T et al. (1999) A model for a umuDC-dependent prokaryotic DNA damage checkpoint. Proc Natl Acad Sci U S A 96:9218-23
Hickman, M J; Samson, L D (1999) Role of DNA mismatch repair and p53 in signaling induction of apoptosis by alkylating agents. Proc Natl Acad Sci U S A 96:10764-9
Li-Sucholeiki, X C; Khrapko, K; Andre, P C et al. (1999) Applications of constant denaturant capillary electrophoresis/high-fidelity polymerase chain reaction to human genetic analysis. Electrophoresis 20:1224-32
Bennett, R A (1999) The Saccharomyces cerevisiae ETH1 gene, an inducible homolog of exonuclease III that provides resistance to DNA-damaging agents and limits spontaneous mutagenesis. Mol Cell Biol 19:1800-9
Ekstrom, P O; Borresen-Dale, A L; Qvist, H et al. (1999) Detection of low-frequency mutations in exon 8 of the TP53 gene by constant denaturant capillary electrophoresis (CDCE). Biotechniques 27:128-34
Glassner, B J; Posnick, L M; Samson, L D (1998) The influence of DNA glycosylases on spontaneous mutation. Mutat Res 400:33-44
Glassner, B J; Rasmussen, L J; Najarian, M T et al. (1998) Generation of a strong mutator phenotype in yeast by imbalanced base excision repair. Proc Natl Acad Sci U S A 95:9997-10002
Masuda, Y; Bennett, R A; Demple, B (1998) Dynamics of the interaction of human apurinic endonuclease (Ape1) with its substrate and product. J Biol Chem 273:30352-9

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