Structural Studies of cullin-based ubiquitin ligases

基于 cullin 的泛素连接酶的结构研究

• 批准号: 7650667
• 负责人: NING ZHENG
• 金额: $ 30.16万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7650667
• 关键词: Adaptor Signaling Protein; Address; Agonist; Antineoplastic Agents; Auxins; BTB/POZ Domain; Biochemical; Biology; Cell Nucleus; Cell physiology; Complex; Cytosol; DNA Repair; DNA biosynthesis; Disease; Docking; Drug Delivery Systems; Enzymes; Eukaryotic Cell; F-Box Proteins; Family; Foundations; Funding; Future; Genome; Hormone Receptor; Hormones; Human; Human Genome; JUN gene; Length; Ligase; Link; Malignant Neoplasms; Mediating; Molecular; Organism; Oxidative Stress; Perception; Physiological; Plant Growth Regulators; Plant Physiology; Plants; Polyubiquitin; Polyubiquitination; Proteins; Proteomics; Publishing; Recruitment Activity; Regulation; Role; Signal Transduction; Structure; System; Ubiquitin; Ubiquitination; Variant; Virus Diseases; base; biological adaptation to stress; cell growth regulation; dimer; human disease; inhibitor/antagonist; jasmonate; nervous system disorder; novel; numb protein; protein function; public health relevance; receptor; response; scaffold; sensor; small molecule; structural biology; transcription factor; tumorigenesis; ubiquitin ligase; ubiquitin-protein ligase

DESCRIPTION (provided by applicant): The cullin-RING ligases (CRLs) represent the largest super-family of multi-subunit E3 ubiquitin ligase complexes in eukaryotic cells. Organized in a modular form with a cullin-RING catalytic scaffold, an adaptor, and an interchangeable substrate-receptor subunit, the CRL1-5 complexes regulate a wide variety of cellular processes by promoting ubiquitination of a large number of protein substrates. Dysregulation and malfunction of CRLs are implicated in tumorigenesis, cancers, and a variety of other human diseases. This project focuses on structure-function studies of several CLR complexes in the regulation of cell growth, DNA replication, DNA repair, oxidative stress response, and hormone perception. In Aim1, a structural biology approach will be used to investigate how the substrate receptor subunits (DCAFs) of a newly identified family of CLRs, the CRL4 E3 complexes, are recruited to the ubiquitin ligase machinery. This study is aimed at resolving the current controversy over the identity of the CRL4 substrate receptors among all WD40-repeat proteins in the human genome. In Aim 2, a combination of biochemical and structural biology approaches will be employed to analyze the structural and functional roles of a novel functional ubiquitin E3 variant protein in the CRL4-Det1 E3 complex, which ubiquitinates the bZIP family of transcription factors such as c-Jun. In Aim 3, the crystal structure of the full length Keap1 dimer will be determined to elucidate how the BTB-domain protein functions as an oxidative stress sensor as well as a dimeric substrate receptor subunit of the CRL3 E3 complex. In Aim 4, the structural mechanism by which the plant COI1 F-box protein functions as the jasmonate plant hormone receptor will be investigated. Together with the recently published results of how another plant F-box protein TIR1 recognizes the plant hormone auxin, this study is aimed at establishing the structural principles of how these naturally occurring small molecules regulate the CRL E3s as agonists and laying the foundation of ongoing and future efforts of developing anti-cancer drugs targeting human CRLs. PUBLIC HEALTH RELEVANCE Tumorigenesis, cancer, and a variety of other human diseases including neurological disorders and viral infections are associated with the abnormal functions of a newly identified family of enzymatic complexes, known as the cullin-RING ubiquitin ligases. This proposal is aimed at studying the structural basis of how these enzyme machineries assemble and function in human and other organisms so that new strategies can be derived to develop novel drugs targeting this disease-associated enzymes.

描述（由申请人提供）：Cullin-Ring连接酶（CRL）代表真核细胞中最大的多亚基E3泛素连接酶复合物的最大超级家庭。 CRL1-5复合物以cullin-ring催化支架，适配器和可互换的底物 - 受体亚基的形式组织，CRL1-5复合物通过促进大量蛋白质底物的泛素化来调节多种细胞过程。 CRL的失调和故障与肿瘤发生，癌症和各种其他人类疾病有关。该项目侧重于在细胞生长，DNA复制，DNA修复，氧化应激反应和激素感知的调节中对几种CLR复合物的结构功能研究。在AIM1中，将使用一种结构生物学方法来研究新鉴定的CLR家族的底物受体亚基（DCAF）如何募集到泛素连接酶机械上。这项研究旨在解决人类基因组中所有WD40重复蛋白中CRL4底物受体身份的当前争议。在AIM 2中，将采用生化和结构生物学方法的组合来分析CRL4-DET1 E3复合物中新型功能性泛素E3变体蛋白的结构和功能作用，从而将BZIP家族泛素化的转录因子（例如C-Jun）（例如C-Jun）。在AIM 3中，将确定全长Keap1二聚体的晶体结构，以阐明BTB域蛋白如何用作氧化应激传感器以及CRL3 E3复合物的二聚体底物受体亚基。在AIM 4中，将研究植物COI1 F-box蛋白作为jasmonate植物激素受体的结构机制。连同最近发表的另一个植物F-box蛋白TIR1如何识别植物激素生长素的结果，该研究旨在建立这些天然存在的小分子如何调节CRL E3的结构原理，并奠定了抗癌症靶向人类CRL的持续和未来努力的基础。公共卫生相关性肿瘤发生，癌症以及包括神经系统疾病和病毒感染在内的各种其他人类疾病与新鉴定的酶复合物家族的异常功能有关，称为库林环泛素连接酶。该建议旨在研究这些酶机如何在人和其他生物中组装和功能的结构性基础，以便可以得出新的策略来开发针对这种疾病相关酶的新型药物。