REGULATION OF PAXILLIN SIGNALING

描述：巴西林是一个70 kDa的蛋白质，定位于局灶性粘连， 细胞与底物的附着点。酪氨酸 另一种粘着斑蛋白p125FAK和巴西林的磷酸化， 受到多种不同的病毒和细胞因子的刺激。近期 申请者作为博士后研究员进行的研究表明， P125FAK和帕西林相互作用，pp125FAK可以磷酸化 帕克西林。拟议研究的中心假设是 Pp125FAK介导的帕西林酪氨酸磷酸化调节 通过募集含有SH2的蛋白质，形成信号复合体。 描述了四个具体目标。Pp125FAK和Paxlin内的序列 哪些是中介蛋白质：蛋白质的相互作用将使用 酵母双杂交法(AIM I)。接下来，src和FAK介导的位点 通过突变酪氨酸来改变帕西林的磷酸化 苯丙氨酸，以及突变的帕西林蛋白结合的能力 将评估已知蛋白质的Sh2结构域(AIM II)。在《目标3》中， 与巴西林结合的蛋白质，已知的和未知的都将被检测。 最后，含有巴西林的信号复合体的重要性将 通过破坏结合在一起的含SH2的蛋白质复合体来确定 酪氨酸磷酸化的巴西林。对细胞转化和细胞生长的影响 将对细胞迁移进行研究。

DESCRIPTION: Paxillin is a 70 kDa protein localized to focal adhesions, points of attachment of the cell to the substratum. Tyrosine phosphorylation of paxillin and p125FAK, another focal adhesion protein, is stimulated by a number of different viral and cellular agents. Recent studies, performed by the applicant as a Postdoctoral Fellow, indicate that p125FAK and paxillin interact, and that pp125FAK can phosphorylate paxillin. The central hypothesis of the proposed research is that pp125FAK mediated tyrosine phosphorylation of paxillin regulates the formation of signaling complexes, by recruiting SH2 containing proteins. Four specific Aims are described. Sequences within pp125FAK and paxillin which mediate protein:protein interaction will be defined using the yeast two-hybrid method (Aim I). Next, the sites of Src and FAK-mediated phosphorylation on paxillin will be altered by mutating tyrosines to phenylalanines, and the ability of the mutated paxillin proteins to bind SH2 domains of known proteins will be assessed (Aim II). In Aim III, proteins that bind to paxillin, both known and unknown will be examined. Finally, the importance of paxillin containing signaling complexes will be determined by disrupting SH2-containing protein complexes bound to tyrosine phosphorylated paxillin. The effects on cell transformation and cell migration will be studied.