S POMBE CAMP SIGNAL PATHWAY

描述：调查员提出以下具体目标：目标 #1：研究腺苷环化酶(GIT2)和腺苷环化酶之间的物理相互作用 直接参与其激活的蛋白质。私家侦探确认了8个 具有正常体外催化活性的改变的git2蛋白(测量 在有Mn++存在的情况下)，但在体内不起作用(称为类别1)。 被测序的4个改变的1类git蛋白的突变 有突变发生在基因的一个小区域内(编码大约 蛋白质的8%)。这些突变可能识别git2的一个区域，该区域 与调节蛋白相互作用。派已经分离出了外源基因 2个不同的1类git2突变体的抑制子(SGT)。建议的私利 实验和分析是合理的，应该确定组件 参与腺苷环化酶的激活。无论这些抑制者 确定激活git2的G蛋白途径的组件仍有待于 下定决心。此外，PI将使用2-混合屏幕来识别 Git2相互作用蛋白。PI有一个很好的二级屏幕，他将在那里 专注于与wt git2相互作用的蛋白质，但不是1类改变的蛋白质 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.