The investigator proposes the following specific aims:
Aim #1 : Study the physical interactions between adenylate cyclase (git2) and proteins directly involved in its activation. The PI has identified 8 altered git2 proteins that have normal in vitro catalytic activity (measured in the presence of Mn++) but are not functional in vivo (termed class 1). The mutations in 4 of the altered class 1 git proteins that were sequenced have mutations occurring within a small region of the gene (encoding about 8% of the protein). These mutations may identify a region of git2 that interacts with regulatory proteins. The PI has isolated extragenic suppressors (sgt) of 2 different class 1 git2 mutants. The PI proposed experiments and analysis are reasonable and should identify components involved in adenylate cyclase activation. Whether these suppressors identify components of the G protein pathway to activate git2 remains to be determined. In addition, the PI will use the 2-hybrid screen to identify git2-interacting proteins. The PI has a good secondary screen where he will concentrate on proteins that interact with wt git2 but not a class 1 altered git2.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM046226-07
Application #
2444793
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1991-07-01
Project End
2000-06-30
Budget Start
1997-07-01
Budget End
1998-06-30
Support Year
7
Fiscal Year
1997
Total Cost
Indirect Cost
Name
Boston College
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
045896339
City
Chestnut Hill
State
MA
Country
United States
Zip Code
02467
Mudge, Dayna K; Yang, Fan; Currie, Brian M et al. (2014) Sck1 negatively regulates Gpa2-mediated glucose signaling in Schizosaccharomyces pombe. Eukaryot Cell 13:202-8
Tong, Kevin; Keller, Thomas; Hoffman, Charles S et al. (2012) Schizosaccharomyces pombe Hat1 (Kat1) is associated with Mis16 and is required for telomeric silencing. Eukaryot Cell 11:1095-103
Ivey, F Douglas; Taglia, Francis X; Yang, Fan et al. (2010) Activated alleles of the Schizosaccharomyces pombe gpa2+ Galpha gene identify residues involved in GDP-GTP exchange. Eukaryot Cell 9:626-33
Roux, Antoine E; Leroux, Alexandre; Alaamery, Manal A et al. (2009) Pro-aging effects of glucose signaling through a G protein-coupled glucose receptor in fission yeast. PLoS Genet 5:e1000408
Kao, Richard S; Morreale, Eric; Wang, Lili et al. (2006) Schizosaccharomyces pombe Git1 is a C2-domain protein required for glucose activation of adenylate cyclase. Genetics 173:49-61
Neely, L A; Hoffman, C S (2000) Protein kinase A and mitogen-activated protein kinase pathways antagonistically regulate fission yeast fbp1 transcription by employing different modes of action at two upstream activation sites. Mol Cell Biol 20:6426-34