Eukaryotic cells initiate a multifaceted response to DNA damage that includes the ubiquitination of key substrates, leading either to their degradation or to the modulation of their activity. Despite the importance of the ubiquitination machinery in facilitating this response, the identities of the ubiquitin ligases involved in this pathway and their key targets are just beginning to be discovered. This proposal seeks to utilize novel, high throughput protein identification technologies developed in the investigator's laboratory to identify the key enzymes and substrates involved in this process. Using a mass spectrometry-based approach, they will analyze the global ubiquitination pattern of proteins of S. cerevisiae, determine how this pattern is altered in response to DNA damaging agents, and identify the enzymes responsible for generating this ubiquitin-mediated DNA damage response. A better understanding of how cells employ the ubiquitination pathway to protect themselves from DNA damage will be critical in developing an understanding of how the inactivation of this process can facilitate tumorigenesis.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21ES012021-01
Application #
6569401
Study Section
Special Emphasis Panel (ZES1-LKB-D (FP))
Program Officer
Thompson, Claudia L
Project Start
2002-09-10
Project End
2004-07-31
Budget Start
2002-09-10
Budget End
2003-07-31
Support Year
1
Fiscal Year
2002
Total Cost
$277,800
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Wohlschlegel, James A; Johnson, Erica S; Reed, Steven I et al. (2006) Improved identification of SUMO attachment sites using C-terminal SUMO mutants and tailored protease digestion strategies. J Proteome Res 5:761-70
Sadygov, Rovshan G; Cociorva, Daniel; Yates 3rd, John R (2004) Large-scale database searching using tandem mass spectra: looking up the answer in the back of the book. Nat Methods 1:195-202
Wohlschlegel, James A; Johnson, Erica S; Reed, Steven I et al. (2004) Global analysis of protein sumoylation in Saccharomyces cerevisiae. J Biol Chem 279:45662-8