In Vivo Interactome and Dynamics of Cullin-Ring Ligases

Cullin 环连接酶的体内相互作用组和动力学

基本信息

批准号：
8489863
负责人：
Lan Huang
金额：
$ 19.25万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-01 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8489863
关键词：
Affect Affinity Affinity Chromatography Binding Biological Preservation Biological Process Biology Cell Cycle Progression Cells Cellular biology Chemicals Complex Cullin Proteins Cytolysis DNA Damage DNA Repair Degradation Pathway Development Disease Drug Targeting Environment Eukaryota Family Family member Fluorescence Freezing Future Health Isotope Labeling Laboratories Lead Life Ligase Link Malignant Neoplasms Mammals Maps Mass Spectrum Analysis Membrane Methods Molecular Molecular Target Pathway interactions Pharmaceutical Preparations Post-Translational Protein Processing Proteasome Inhibition Proteins Regulation Signal Transduction Specificity System Technology Therapeutic Tumor Suppressor Genes UV induced DNA damage Ubiquitin Ubiquitination Urea base cancer therapy crosslink drug development extracellular human disease improved in vivo insight multicatalytic endopeptidase complex novel protein complex protein degradation protein protein interaction protein purification receptor response therapeutic target ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): Cullin-RING ligases (CRLs) represent the largest family of E3 ubiquitin ligases in eukaryotes and they are involved in regulating a wide array of biological processes including development, cell cycle progression, signal transduction, and DNA damage. CRLs are key players in the ubiquitin-proteasome system (UPS) and control protein fate by marking specific substrates for ubiquitination and subsequent proteasomal degradation. Unlike other RING E3 ligases, cullins do not bind substrates directly; instead, CRL specificity relies mainly on substrate receptors that dynamically interact with core CRL complexes through protein-protein interactions. In mammals, there are multiple cullin family members and specific adaptors, as well as a large number of substrate receptors, which can assemble in a modular manner to form over 400 CRLs in vivo. CRLs are estimated to target nearly 20% of proteasomal degradation substrates including many oncogenes and tumor suppressor genes. Given its critical importance in the UPS and cell biology, dis-regulation of CRLs' function can lead to many different diseases including cancer. Therefore, detailed understanding of the CRL biology would not only further our understanding of how they are regulated during cancer development but also provide novel molecular targets for future cancer therapeutics targeting at selective protein degradation. This would be a much more attractive strategy for developing more effective and less toxic cancer treatment in comparison to current drugs targeting the UPS through general proteasome inhibition. Protein-protein interactions are essential for establishing CRLs' diverse assemblies and activities. We hypothesize that mapping the CRL interaction networks in vivo not only would allow us to obtain a complete description of CRL system in living cells but also has important implications for the identification of pharmacological agents that affect particular CRL pathways. To obtain the CRL landscape in living cells, we propose to employ the QTAX strategy to define in vivo protein interactome and dynamics of CRLs, aiming to identify new regulators of the CRL complexes. Here are our specific aims: 1) To define the in vivo interactome of CRL complexes as they occur in living cells; 2) To unravel the in vivo interaction dynamics of CRL4 complexes during DNA damage response to elucidate their function and regulation.

描述（由申请人提供）：Cullin-RING 连接酶 (CRL) 代表真核生物中最大的 E3 泛素连接酶家族，它们参与调节多种生物过程，包括发育、细胞周期进程、信号转导和 DNA 损伤。 CRL 是泛素蛋白酶体系统 (UPS) 的关键参与者，通过标记泛素化和随后的蛋白酶体降解的特定底物来控制蛋白质的命运。与其他 RING E3 连接酶不同，cullin 不直接结合底物；相反，CRL 特异性主要依赖于通过蛋白质-蛋白质相互作用与核心 CRL 复合物动态相互作用的底物受体。哺乳动物体内存在多个cullin家族成员和特异性接头，以及大量的底物受体，它们可以在体内以模块化方式组装形成400多个CRL。据估计，CRL 靶向近 20% 的蛋白酶体降解底物，包括许多癌基因和抑癌基因。鉴于其在 UPS 和细胞生物学中的至关重要性，CRL 功能的失调可能导致包括癌症在内的许多不同疾病。因此，对 CRL 生物学的详细了解不仅可以加深我们对它们在癌症发展过程中如何调节的理解，而且还可以为未来针对选择性蛋白质降解的癌症治疗提供新的分子靶点。与目前通过一般蛋白酶体抑制来靶向 UPS 的药物相比，这将是开发更有效且毒性更低的癌症治疗方法的更有吸引力的策略。蛋白质-蛋白质相互作用对于建立 CRL 的多样化组装和活性至关重要。我们假设，绘制体内 CRL 相互作用网络不仅能让我们获得活细胞中 CRL 系统的完整描述，而且对识别影响特定 CRL 途径的药物制剂具有重要意义。为了获得活细胞中的 CRL 景观，我们建议采用 QTAX 策略来定义体内蛋白质相互作用组和 CRL 动力学，旨在识别 CRL 复合物的新调节因子。以下是我们的具体目标： 1) 定义 CRL 复合物在活细胞中发生的体内相互作用组； 2) 揭示DNA损伤反应过程中CRL4复合物的体内相互作用动力学，以阐明其功能和调节。