There are 3 ways in which mismatched bases arise in DNA: 1) Misincorporation during DNA replication; 2) Production of regions of heteroduplex DNA during genetic recombination; And 3) chemical damage to DNA and DNA precursors. Mismatch repair (MMR) limits recombination between related DNA sequences containing base differences or between short repeated sequences thus reducing the frequency of aberrant recombination events. Failure to repair mispaired bases increases the spontaneous mutation rate and also gives rise to altered recombination events. Understanding the mechanism of MMR will impact human health for a number of reasons: 1) Hereditary nonpolyposis colon cancer is due to inherited defects in MMR and many sporadic cancers appear MMR defective yet not all of the genes that underlie these diseases are known; 2) Many chemotherapy agents damage DNA and understanding MMR could lead to improvements in the efficacy of these agents; And 3) purification of MMR proteins will provide new reagents for detecting base changes in DNA which will be useful for genetic studies. The goal of this proposal is to identify Saccharomyces cerevisiae MMR proteins and understand how they catalyze MMR. Associated goals are to understand how MMR interacts with genetic recombination, how MMR contributes to the fidelity of DNA replication and if defects in MMR are responsible for inherited diseases. The following lines of experimentation will be carried out: 1) Genetic studies will identify MMR genes and help understand how they function in MMR; 2) Two-hybrid interaction analysis will be continued to identify and study genes encoding proteins that interact with MSH2, MSH3 and MSH6; 3) Biochemical characterization of an in vitro MMR system will be continued to identify and purify enzymes required for MMR; 4) Biochemical studies of individual MMR proteins including the MSH2-MSH3, MSH2-MSH6, MLH1-PMS1 and MLH1-MLH3 complexes, RPA, PCNA and EXO1 will be continued to determine the roles these proteins play in MMR. The ultimate goal of these experiments is to reconstitute MMR with purified proteins and determine the mechanism of this reaction. It is also anticipated that these studies will provide tools for identifying additional MMR genes in higher eukaryotes for use in studying MMR and the genetics of human cancer susceptibility.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM050006-10
Application #
2752342
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1988-07-01
Project End
2002-11-30
Budget Start
1998-12-01
Budget End
1999-11-30
Support Year
10
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of California San Diego
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
077758407
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Perrella, Giorgio; Davidson, Mhairi L H; O'Donnell, Liz et al. (2018) ZINC-FINGER interactions mediate transcriptional regulation of hypocotyl growth in Arabidopsis. Proc Natl Acad Sci U S A 115:E4503-E4511
Graham 5th, William J; Putnam, Christopher D; Kolodner, Richard D (2018) The properties of Msh2-Msh6 ATP binding mutants suggest a signal amplification mechanism in DNA mismatch repair. J Biol Chem 293:18055-18070
Bowen, Nikki; Kolodner, Richard D (2017) Reconstitution of Saccharomyces cerevisiae DNA polymerase ?-dependent mismatch repair with purified proteins. Proc Natl Acad Sci U S A 114:3607-3612
Huang, He; Alvarez, Sophie; Bindbeutel, Rebecca et al. (2016) Identification of Evening Complex Associated Proteins in Arabidopsis by Affinity Purification and Mass Spectrometry. Mol Cell Proteomics 15:201-17
Kolodner, Richard D (2016) A personal historical view of DNA mismatch repair with an emphasis on eukaryotic DNA mismatch repair. DNA Repair (Amst) 38:3-13
Putnam, Christopher D (2016) Evolution of the methyl directed mismatch repair system in Escherichia coli. DNA Repair (Amst) 38:32-41
Reyes, Gloria X; Schmidt, Tobias T; Kolodner, Richard D et al. (2015) New insights into the mechanism of DNA mismatch repair. Chromosoma 124:443-62
Smith, Catherine E; Bowen, Nikki; Graham 5th, William J et al. (2015) Activation of Saccharomyces cerevisiae Mlh1-Pms1 Endonuclease in a Reconstituted Mismatch Repair System. J Biol Chem 290:21580-90
Kaiserli, Eirini; Páldi, Katalin; O'Donnell, Liz et al. (2015) Integration of Light and Photoperiodic Signaling in Transcriptional Nuclear Foci. Dev Cell 35:311-21
Goellner, Eva M; Putnam, Christopher D; Kolodner, Richard D (2015) Exonuclease 1-dependent and independent mismatch repair. DNA Repair (Amst) 32:24-32

Showing the most recent 10 out of 91 publications