ROLE OF TRIP6 IN P130CAS SIGNALING AND CELL MOTILITY

信号分子通过衔接子或支架蛋白定位于离散的细胞微环境正在成为解释信号转导级联的效率和特异性的范例。 因此，参与组织这些“信号体”的蛋白质在细胞内通讯的调节中起着关键作用。 TRIP 6 cDNA编码具有富含脯氨酸的区域和三个连续的LIM结构域的蛋白质。富含脯氨酸的区域具有与Src-同源性-3（SH 3）结构域相互作用的能力，并且LIM结构域也被表征为蛋白质/蛋白质相互作用基序。 Beckerle博士实验室的证据表明TRIP 6与另一种粘着斑分子p130 cas之间存在关联。 p130 cas是一种非酶信号蛋白，其在响应整合素刺激的粘附和细胞迁移时被Src和粘着斑激酶酪氨酸磷酸化。 理解这些现象的努力是重要的，因为异常的粘附和迁移可导致病理生理学，如癌症。根据这些观察，我建议TRIP 6作为一个衔接蛋白的功能作用，以协调参与整合素介导的粘附，如p130 cas和其他尚未确定的蛋白质的信号转导。 该提案的目标计划确定TRIP 6在调节p130 cas信号传导和细胞运动性中的相关性。 本研究的目的是：1）定位TRIP 6和p130 cas的结合结构域; 2）评估破坏p130 cas/TRIP 6相互作用的影响; 3）鉴定新的TRIP 6结合伴侣。

The localization of signalling molecules to discrete cellular microenvironments via adaptor or scaffolding proteins is emerging as a paradigm to explain the efficiency and specificity of signal transduction cascades. Thus, proteins involved in organizing these "signalsomes" play a pivotal role in the regulation of intracellular communication. The TRIP6 cDNA encodes a protein with a proline-rich region and three consecutive LIM domains Proline-rich regions have the capacity to interact with Src- homology-3 (SH3) domains and LIM domains also have been characterized as protein/protein interaction motifs. Evidence in Dr. Beckerle's laboratory indicates an association between TRIP6 and another focal adhesion molecule, p130cas. p130cas is a nonenzymatic signalling protein that is tyrosine phosphorylated by Src and focal adhesion kinase in response to integrin- stimulated adhesion and cell migration. Efforts to understand these phenomenon are important because aberrant adhesion and migration can contribute to pathologic physiology such as cancer. In light of these observations, I propose a functional role for TRIP6 as an adaptor protein to coordinate signal transducers involved in integrin-mediated adhesion such as p130cas and other as of yet unidentified proteins. The goal of this proposal plans to determine the relevance of TRIP6 in the regulation of p130cas signalling and cell motility. The aims of this proposal are to 1) map the-binding domains on TRIP6 and p130cas; 2)assess the effects of disrupting the p130cas/TRIP6 interaction, and 3) identify new TRIP6 binding partners.