Phosphatase mediated regulation of PKC and Akt

描述（由申请人提供）： 本研究的长期目标是阐明蛋白磷酸酶介导的蛋白激酶C和Akt调节的分子机制。 蛋白激酶C和Akt属于AGC激酶超家族。 这两种激酶已被证明在调节肿瘤发生过程中发挥关键作用。 这两种激酶都受多步磷酸化过程的调节。 虽然激酶的磷酸化对酶活性很重要，但去磷酸化与酶失活和下调有关。 然而，对蛋白激酶C和Akt的去磷酸化过程知之甚少。 我们最近发现了一种新的pleckstrin同源蛋白磷酸酶，SCOP。 初步研究表明，这种磷酸酶在体外和体内使蛋白激酶C和Akt去磷酸化。 该提案解决了SCOP如何调节蛋白激酶C和Akt。 具体目标是： 1)SCOP介导的体内去磷酸化对PKC和Akt的调节：本节的目的是了解SCOP使细胞中PKC和Akt去磷酸化的分子机制。 具体而言，SCOP的活性是否需要膜结合和脂质信号传导将被研究。 2)SCOP介导的去磷酸化的功能意义：使用siRNA方法敲低内源性SCOP的表达水平，并研究蛋白激酶C和Akt的功能变化。 3)通过蛋白质：蛋白质相互作用调节SCOP：本节的目的是了解SCOP与蛋白激酶C或Akt之间以及SCOP与其他细胞蛋白之间的蛋白质：蛋白质相互作用如何影响SCOP功能。 我的长期职业目标是成为癌症生物学研究领域的独立科学调查员。 具体来说，我对研究癌症的分子机制感兴趣。 为了实现这个目标，我的短期计划是在一个顶级学术机构的一个完善的实验室获得更多的研究经验。 牛顿博士在加州大学圣地亚哥分校的实验室将为我提供进一步发展我的职业生涯所需的额外培训。 有了这笔资助，我将有机会独立工作，制定自己的研究计划。 同时，我将从牛顿博士和UCSD提供的优秀研究设施中受益匪浅，并从与UCSD教职员工的科学交流中受益匪浅。

DESCRIPTION (provided by applicant): The long term goal of the proposed research is to elucidate the molecular mechanism of protein phosphatase mediated regulation of protein kinase C and Akt. Protein kinase C and Akt belong to the AGC kinase superfamily. Both kinases have been shown to play critical roles in modulating tumorigenesis process. Both kinases are regulated by a multi-step phosphorylation process. While phosphorylation of the kinases is important for the enzyme activity, dephosphorylation is associated with enzyme inactivation and downregulation. However, little is known about the dephosphorylation process of protein kinase C and Akt. We have recently identified a novel pleckstrin homology containing protein phosphatase, SCOP. Preliminary studies have indicated that this phosphatase dephosphorylates protein kinase C and Akt both in vitro and in vivo. This proposal addresses how SCOP regulates protein kinase C and Akt. Specific Aims are: 1) Regulation of PKC and Akt by SCOP-mediated dephosphorylation in vivo: The goal of this section is to understand the molecular mechanism by which SCOP dephosphorylates PKC and Akt in cells. Specifically, whether the activity of SCOP requires membrane binding and lipid signaling will be investigated. 2) Functional significance of SCOP-mediated dephosphorylation: Expression level of endogenous SCOP will be knocked down using siRNA approach, and functional changes of protein kinase C and Akt will be studied. 3) Regulation of SCOP by protein:protein interactions: The goal of this section is to understand how protein:protein interactions between SCOP and protein kinase C or Akt, and between SCOP and other cellular proteins affect SCOP function. My long-term career goal is to become an independent scientific investigator in the field of cancer biology research. Specifically, I am interested in studying the molecular mechanisms of cancer. To achieve this goal, my short-term plan is to gain more research experience in a well established laboratory at a top academic institute. Dr. Newton's lab at UCSD will provide me with the additional training that I need to develop my career further. With the support by this grant, I will have the opportunity to work independently and develop my own research plan. Meanwhile, I will be greatly benefited from the outstanding research facilities provided by Dr. Newton and UCSD, and from scientific communications with the faculty members at UCSD.