The objectives of this research are to discover the proliferating cell nuclear antigen (PCNA) binding site on yeast MutLalpha, a component of the mismatch repair machinery, and to discover if MutLalpha can interact with the 9-1-1 complex, a set of proteins involved in cell cycle checkpoint control. In addition to mismatch repair, MutLalpha is involved in apoptosis signaling, nucleotide excision repair, and meiotic recombination. Mutator phenotypes of variant MutLalpha complexes include cancer and infertility. This research would determine how and if PCNA and/or the 9-1-1 complex are also involved in these processes. These findings would clearly have important implications for cancer and fertility research. To reach the objectives of the project, protein mutagenesis will be used to make pre-determined variant MutLalpha complexes. Both wild-type and variant complexes will be assayed for their interaction with PCNA (and the 9-1-1 complex) in vitro using affinity chromatography and atomic force microscopy and in vivo using a yeast two-hybrid assay. Variants which do not interact with PCNA (or the 9-1-1 complex) will be assayed for their mutator phenotypes.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32GM069100-03
Application #
6895215
Study Section
Special Emphasis Panel (ZRG1-F08 (20))
Program Officer
Portnoy, Matthew
Project Start
2003-07-01
Project End
2006-06-30
Budget Start
2005-07-01
Budget End
2006-06-30
Support Year
3
Fiscal Year
2005
Total Cost
$47,196
Indirect Cost
Name
U.S. National Institute of Environ Health Sciences
Department
Type
DUNS #
040034860
City
Research Triangle Park
State
NC
Country
United States
Zip Code
27709
Holmes, Shannon F; Scarpinato, Karin Drotschmann; McCulloch, Scott D et al. (2007) Specialized mismatch repair function of Glu339 in the Phe-X-Glu motif of yeast Msh6. DNA Repair (Amst) 6:293-303