SSK2 ASSOCIATED PROTEINS

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。MEK激酶Ssk2p是人类MEK激酶MTK1/MEKK4的同源物，具有调节肌动蛋白细胞骨架以响应渗透胁迫的独特性质。我们的工作主要集中在酵母中，即酵母激酶Ssk2p，但我们之前已经证明，人类激酶具有这些独特的能力。从基因实验中我们知道，该激酶的调节独立于其正常的MAPK通路，它直接与肌动蛋白结合，在所有酵母中，MEK激酶具有这种独特的调节肌动蛋白组装的能力。这项合作是为了尝试并确定Ssk2p相关蛋白，这些蛋白可能负责途径无关的激酶调节，并可能是该激酶的细胞骨架效应因子(S)。方法是对Ssk2p进行TAP标记，从酵母中沉淀复合体，并使用质谱学来鉴定这些相关蛋白质。

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. The MEK kinase Ssk2p, a homolog of the human MEK kinase MTK1/MEKK4 has unique properties for this class of kinases in being able to regulate the actin cytoskeleton in response to osmotic stress. Our work is focused in yeast, with the yeast kinase Ssk2p, but we have previously shown that the human kinase has these unique capabilities. We know from genetic experiments that this kinase is regulated independently of its normal MAPK pathway, that it binds actin directly and among all yeast MEK kinases has this unique capacity to regulate actin assembly. This collaboration was initiated to try and identify Ssk2p-associated proteins that could be responsible for pathway independent kinase regulation and that could be the cytoskeletal effector(s) of the kinase. The approach is to TAP-tag Ssk2p, precipitate complexes from yeast and use mass-spec to identify these associated proteins.