S POMBE CAMP SIGNAL PATHWAY

描述：研究者提出以下具体目标： #1：研究腺苷酸环化酶（git 2）和 直接参与其激活的蛋白质。 PI已确定8个 具有正常体外催化活性的改变的Git 2蛋白（测量 在Mn++存在下），但在体内不起作用（称为1类）。 测序的4个改变的1类git蛋白中的突变 在基因的一个小区域内发生突变（编码约 8%的蛋白质）。 这些突变可以识别git 2的一个区域， 与调节蛋白相互作用。 私家侦探已经分离出了 2种不同的1类GIT 2突变体的抑制子（SGT）。 PI建议 实验和分析是合理的， 参与腺苷酸环化酶的激活。 这些抑制剂 确定G蛋白通路的组分以激活git 2仍然是 测定 此外，PI将使用2-混合筛选来识别 git 2相互作用蛋白。 PI有一个很好的辅助屏幕，他将 集中于与野生型git 2相互作用但不改变1类的蛋白质 git2。

DESCRIPTION: 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.