CHARACTERIZATION OF SRC AS AN E6AP SUBSTRATE

描述 泛素介导的蛋白质水解是蛋白质降解的主要机制之一。 调节细胞信号网络的蛋白质组成。 E6 AP最初在Howley实验室被鉴定为人类 乳头瘤病毒（HPV）16 E6结合蛋白，作为一种泛素- 蛋白连接酶（E3）在p53降解中的作用。在不存在HPV 16 E6、E6 AP的情况下， 已知作为E3酶发挥作用，然而，其底物仍然存在 不清楚豪利实验室最近的工作已经确定了几个 E6 AP的潜在底物，包括Src家族成员Blk 非受体酪氨酸激酶。E6 AP对BLK作用的研究 已经证明E6 AP选择性靶向激酶活性形式 用于降解。本提案的目的是确定， E6 AP影响c-Src和V-Src的稳定性。初步数据 表明E6 AP与c-Src相互作用实际上是E6 AP的底物 以及这种激酶的活化形式是否被选择性降解。在 此外，将研究E6 AP对v-Src的影响，以确定是否 v-被选择性降解。此外，E6 AP对v-SRC的影响也不明显。 将进行研究，以确定v-SRC是否不稳定或能够逃逸 定向降解。HPV 16 E6对E6 AP介导的细胞凋亡的影响 由于HPV 16 E6可能改变c-Src的表达， 由于E6 AP的底物特异性，c-Src的降解也将被 分析，因为HPV 16 E6可以改变E6 AP的底物特异性。 这些研究将进一步加深我们对人类癌症的理解 通过提供对细胞转化的洞察， 酪氨酸激酶和人乳头瘤病毒。

DESCRIPTION Ubiquitin-mediated proteolysis is one of the major mechanisms used by the cell to regulate the protein composition of signaling networks. E6AP, which was originally identified in the Howley lab as a human papillomavirus (HPV) 16 E6 binding protein, functions as a ubiquitin- protein ligase (E3) in p53 degradation. In the absence of HPV16 E6, E6AP is known to function as an E3 enzyme, however, its substrates remain unclear. Recent work from the Howley lab has identified several potential substrates for E6AP, including Blk, a member of the Src family of non-receptor tyrosine kinases. Studies of the effects of E6AP on BLK have demonstrated that E6AP selectively targets the kinase active form of Blk for degradation. The goal of this proposal is to determine if E6AP affects the stability of c-Src and V-Src. Preliminary data indicates that E6AP interacts with c-Src is in fact a substrate for E6AP and if the activated forms of this kinase are selectively degraded. In addition, the effects of E6AP on v-Src will be studied to determine if v- are selectively degraded. In addition, the effects of E6AP on v-SRC will be studied to determine if v-SRC is unstable or is able to escape targeted degradation. The effects of HPV16 E6 on E6AP mediated degradation of c-Src will also be analyzed, as HPV16 E6 may alter he substrate specificity of E6AP, degradation of c-Src will also be analyzed, as HPV16 E6 may alter the substrate specificity of E6AP. Together, these studies will further our understanding of human cancer by providing insight into cellular transformation both by non-receptor tyrosine kinases and by human papillomaviruses.