This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.Our lab has identified mutations in several genes required for adenylate cyclase activation in fission yeast. These genes encode the subunits of a heterotrimeric G protein and a likely GPCR, along with three genes not previously known to be associated with cAMP signaling. We also have a collection of mutations in the adenylate cyclase gene that appear to affect activation but not catalytic activity. We would like to complement our genetic analyses with a study of the proteins physically associated with adenylate cyclase in fission yeast as a function of the carbon source environment. We would also like to study the effect of mutations within adenylate cyclase on these interactions. We have constructed an integrated TAP-tagged form of adenylate cyclase at its endogenous locus and are able to detect it by Western analysis. We are now using the 'Purification of TAP-Tagged Proteins' protocol from the Yeast Resource Center to allow us to isolate protein complexes from this strain.
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