ADENYLATE CYCLASE-ASSOCIATED COMPLEXES IN SCHIZOSACCHAROMYCES POMBE

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 我们的实验室已经确定了几个基因的突变所需的腺苷酸环化酶激活裂变酵母。这些基因编码异源三聚体G蛋白的亚基和可能的GPCR，沿着三个先前不知道与cAMP信号传导相关的基因。我们还收集了腺苷酸环化酶基因的突变，这些突变似乎影响激活，但不影响催化活性。我们想补充我们的遗传分析与研究的蛋白质物理相关的腺苷酸环化酶在裂变酵母作为一个功能的碳源环境。我们还想研究腺苷酸环化酶内的突变对这些相互作用的影响。我们已经构建了一个整合的TAP标记形式的腺苷酸环化酶在其内源性位点，并能够检测到它的Western分析。我们现在正在使用酵母资源中心的“TAP标记蛋白质的纯化”方案，使我们能够从该菌株中分离蛋白质复合物。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for 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.