SPECIFICITY OF UBIQUITINATION--THE HPV E6/E6 AP PARADIGM

泛素化的特异性——HPV E6/E6 AP 范式

• 批准号: 2010998
• 负责人: JON HUIBREGTSE
• 金额: $ 21.4万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-12-13 至 2001-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2010998
• 关键词: Saccharomyces cerevisiae; acid aminoacid ligase; enzyme activity; human papillomavirus; microorganism culture; oligonucleotides; site directed mutagenesis; tumor suppressor proteins; ubiquitin; virus genetics; virus protein

DESCRIPTION: E6-AP (E6-Associated Protein) is a cellular protein that associates with and catalyzes ubiquitination of p53 in anE6-dependent manner. Characterization of E6-AP showed it to be representative of a large family of ubiquitin-protein ligases (or E3 proteins), which have been postulated to play the major role in substrate selectivity. The applicant proposes a model for function of the E6-AP-related E3 proteins which states that they consist of two broad domains: an N-terminal substrate recognition domain, and a C-terminal catalytic domain (the hect domain, for homologous to E6-AP C-terminus) which ubiquitinates bound substrates. The specific aims of this proposal are: 1) to evaluate this model by determining if substrate specificity can be redirected by substitution of N-terminal sequences and if hect domains are functionally interchangeable among E3 proteins, 2) to evaluate this model with respect to E6-AP by precisely mapping the determinants necessary for p53 association and hect domain function, and 3) to identify substrates of Rsp5, an essential yeast hectE3, beginning with predictions of the 2-domain model and taking a combined genetic and biochemical approach. The long-term objectives of this application are to understand the mechanisms of action of the hect E3 proteins and their role, as well as the role of the high-risk HPV E6 proteins (and possible cellular analogs), in directing the specificity of the ubiquitin system. The ultimate goal is a more complete understanding of HPV-associated carcinogenesis and other processes regulated by protein ubiquitination.

描述：E6-AP（与E6相关蛋白）是一种细胞蛋白 与Ane6依赖性中p53的并催化并催化泛素化 方式。 E6-AP的表征表明它代表了大型 泛素蛋白蛋白连接酶（或E3蛋白）的家族， 假设在底物选择性中起主要作用。 申请人 提出了一个指出E6-AP相关的E3蛋白功能的模型 它们由两个广泛的领域组成：N末端底物识别 域和一个C末端催化域（用于同源的​​HECT结构域 到E6-AP C末端），泛素化结合的底物。 具体 该提案的目的是：1）通过确定是否是否确定该模型 底物特异性可以通过替代N端重定向 序列以及HECT域在E3之间的功能上可以互换 蛋白质，2）通过精确评估有关E6-AP的该模型 绘制p53关联所需的决定因素并缩小域 功能，3）识别RSP5的底物，必需的酵母Hote3， 首先要预测2域模型，然后进行组合 遗传和生化方法。 这个长期目标 应用是要了解E3 hect hect的作用机理 蛋白质及其作用以及高风险HPV E6的作用 蛋白质（以及可能的细胞类似物），指导特异性 泛素系统。 最终目标是对 HPV相关的致癌作用和其他由蛋白质调节的过程 泛素化。