The Cullin-ROC Family of E3 Ubiquitin Ligases

E3 泛素连接酶的 Cullin-ROC 家族

• 批准号: 8085407
• 负责人: YUE XIONG
• 金额: $ 9.1万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8085407
• 关键词: Back; Binding; Biochemical; CDT1 Gene; CUL3 gene; CUL4A gene; Cell Proliferation; Cell physiology; Computer Simulation; DNA Damage; Family; Genetic; Genetic Transcription; Goals; Investigation; Knockout Mice; Licensing Factor; Ligase; Mitosis; Mitotic; Mutant Strains Mice; Play; Protein Binding; Proteins; Proteomics; RBX1 gene; Recruitment Activity; Regulation; Replication Licensing; Research; Research Personnel; Role; Series; Transcriptional Regulation; Zinc Fingers; base; in vivo; numb protein; response; ubiquitin ligase; ubiquitin-protein ligase

DESCRIPTION (provided by applicant): This is the first competitive renewal of GM067113, an investigation aimed at the long term goal of understanding the function and regulation of cullin-RING family of E3 ubiquitin ligases (CRLs). Looking back at the journey of the past several years, we are very pleased to see the contribution we made and more remarkably, the advancements achieved collectively by colleagues. Following the initial discovery of ROC1 and ROC2, we have made several findings toward this goal. These include: (1) that CAND1, an evolutionary conserved cullin-associated and neddylation dissociated protein, controls substrate recruitment to all CRLs, (2) that CUL3 binds to a BTB motif present in more than 200 mammalian proteins and potentially assembles a large number of BTB-CUL3-ROC1 ligases, (3) that DDB1-CUL4-ROC1 ligase target the replication licensing factor CDT1 for degradation in response to DNA damage, and (4) that CUL4 interacts with large number of cellular proteins, and (5) that pl70(KIAA080), an evolutionarily-conserved WD40 protein, binds abundantly with CUL4A via DDB1 and plays an essential role in cellular proliferation in a p53-dependent manner. With these series of findings as the basis, we propose a further research to elucidate the substrate- recruitment mechanism, the cellular function and regulation of CUL3- and CUL4-dependent ubiquitin ligases. The studies outlined in this proposal combine computer modeling, proteomic, biochemical, cellular and genetic approaches, offering an excellent and unique opportunity to help in the understanding of the largest family of E3 ubiquitin ligases. Three specific aims are: (i) to determine the functions and substrates of CRLS in mitosis and in transcriptional regulation, (ii) to determine the substrate recruiting mechanism of CRL4 ligases, and (iii) to determine the function and mechanism of p170.

描述（由申请人提供）：这是 GM067113 的首次竞争性更新，这项研究旨在了解 E3 泛素连接酶 (CRL) cullin-RING 家族的功能和调节的长期目标。回顾过去几年的历程，我们很高兴看到我们所做出的贡献，更令人惊叹的是同事们共同取得的进步。继 ROC1 和 ROC2 的初步发现之后，我们为实现这一目标取得了多项发现。其中包括：(1) CAND1，一种进化上保守的 cullin 相关和 neddylation 解离蛋白，控制所有 CRL 的底物招募，(2) CUL3 与 200 多种哺乳动物蛋白中存在的 BTB 基序结合，并可能组装大量蛋白BTB-CUL3-ROC1 连接酶，(3) DDB1-CUL4-ROC1 连接酶靶向复制许可因子 CDT1响应 DNA 损伤而降解，(4) CUL4 与大量细胞蛋白相互作用，(5) pl70(KIAA080)，一种进化上保守的 WD40 蛋白，通过 DDB1 与 CUL4A 大量结合，并发挥重要作用以 p53 依赖性方式参与细胞增殖。以这一系列的发现为基础，我们提出进一步的研究来阐明CUL3和CUL4依赖性泛素连接酶的底物招募机制、细胞功能和调节。该提案中概述的研究结合了计算机建模、蛋白质组学、生物化学、细胞和遗传方法，为帮助了解最大的 E3 泛素连接酶家族提供了绝佳且独特的机会。三个具体目标是：(i) 确定 CRLS 在有丝分裂和转录调控中的功能和底物，(ii) 确定 CRL4 连接酶的底物募集机制，以及 (iii) 确定 p170 的功能和机制。