Staudinger J, Zhou J, Burgess R, Elledge S J, Olson E N
Department of Biochemistry and Molecular Biology, University of Texas M.D. Anderson Cancer Center, Houston 77030.
J Cell Biol. 1995 Feb;128(3):263-71. doi: 10.1083/jcb.128.3.263.
Protein kinase C (PKC) plays a central role in the control of proliferation and differentiation of a wide range of cell types by mediating the signal transduction response to hormones and growth factors. Upon activation by diacylglycerol, PKC translocates to different subcellular sites where it phosphorylates numerous proteins, most of which are unidentified. We used the yeast two-hybrid system to identify proteins that interact with activated PKC alpha. Using the catalytic region of PKC fused to the DNA binding domain of yeast GAL4 as "bait" to screen a mouse T cell cDNA library in which cDNA was fused to the GAL4 activation domain, we cloned several novel proteins that interact with C-kinase (PICKs). One of these proteins, designated PICK1, interacts specifically with the catalytic domain of PKC and is an efficient substrate for phosphorylation by PKC in vitro and in vivo. PICK1 is localized to the perinuclear region and is phosphorylated in response to PKC activation. PICK1 and other PICKs may play important roles in mediating the actions of PKC.
蛋白激酶C（PKC）通过介导对激素和生长因子的信号转导反应，在多种细胞类型的增殖和分化控制中发挥核心作用。在被二酰基甘油激活后，PKC转位至不同的亚细胞位点，在那里它使众多蛋白质磷酸化，其中大多数蛋白质的身份尚不清楚。我们使用酵母双杂交系统来鉴定与活化的PKCα相互作用的蛋白质。利用与酵母GAL4的DNA结合结构域融合的PKC催化区域作为“诱饵”，筛选其中cDNA与GAL4激活结构域融合的小鼠T细胞cDNA文库，我们克隆了几种与C激酶相互作用的新蛋白质（PICKs）。这些蛋白质之一，命名为PICK1，特异性地与PKC的催化结构域相互作用，并且在体外和体内都是PKC磷酸化的有效底物。PICK1定位于核周区域，并响应PKC激活而被磷酸化。PICK1和其他PICKs可能在介导PKC的作用中发挥重要作用。
