ADENYLATE CYCLASE-ASSOCIATE COMPLEXES IN SCHIZOSACCHAROMYCES POMBE

裂殖糖酵母中的腺苷酸环化酶相关复合物

• 批准号: 7420704
• 负责人: CHARLES S. HOFFMAN
• 金额: $ 0.29万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-20 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7420704
• 关键词: adenylate cyclase; gene mutation; genes; proteins; yeasts

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.

该子项目是利用 NIH/NCRR 资助的中心拨款提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中得到体现。列出的机构是中心的机构，不一定是研究者的机构。我们的实验室已经确定了裂殖酵母中腺苷酸环化酶激活所需的几个基因的突变。这些基因编码异源三聚体 G 蛋白的亚基和可能的 GPCR，以及之前未知的与 cAMP 信号传导相关的三个基因。我们还收集了腺苷酸环化酶基因中的一系列突变，这些突变似乎影响激活但不影响催化活性。我们希望通过对裂殖酵母中与腺苷酸环化酶物理相关的蛋白质作为碳源环境的函数的研究来补充我们的遗传分析。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。我们在内源基因座构建了腺苷酸环化酶的集成 TAP 标记形式，并能够通过蛋白质印迹分析对其进行检测。我们现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。