Role of CSN in the activity and dynamic cycling of cullin-RING ubiquitin ligases

CSN 在 cullin-RING 泛素连接酶活性和动态循环中的作用

• 批准号: 8438071
• 负责人: SETH M COHEN
• 金额: $ 34.44万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8438071
• 关键词: Active Sites; Affinity Chromatography; Antineoplastic Agents; Architecture; Behavior; Biochemical; Biological; Biological Assay; Biology; Cancer Etiology; Cancer Model; Cell Proliferation; Cell physiology; Cells; Chemicals; Chemotherapy-Oncologic Procedure; Clinical Trials; Complex; Cullin Proteins; Data; Development; Digit structure; Discrimination; Dominant-Negative Mutation; Drug Targeting; Enzymes; Event; Excision; Exhibits; Family; Fission Yeast; Foundations; Future; Genetic; Guanine Nucleotide Exchange Factors; Histones; Human; IL27RA gene; Inhibitory Concentration 50; Isotope Labeling; KRAS2 gene; Knowledge; Link; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Metalloproteases; Methods; Modification; Molecular Bank; Molecular Genetics; Monitor; Multiple Myeloma; Mutation; Normal Cell; Oncogenic; Pathogenesis; Pathway interactions; Pharmaceutical Preparations; Physiological Processes; Play; Population; Positioning Attribute; Post-Translational Protein Processing; Proteins; Proteome; Proteomics; Protocols documentation; Publishing; Reagent; Regulation; Research; Role; Structure-Activity Relationship; Testing; Thalidomide; Tumorigenicity; Ubiquitin Like Proteins; United States National Institutes of Health; Work; Zinc; cancer cell; cancer therapy; carbonate dehydratase; human disease; inhibitor/antagonist; isopeptidase; lenalidomide; novel; preclinical study; preference; public health relevance; receptor; repository; research study; response; scaffold; small molecule; tool; ubiquitin ligase

DESCRIPTION (provided by applicant): The cullin-RING ubiquitin ligase (CRL) family contains upwards of 240 ubiquitin ligases that regulate a broad range of cellular and organismal processes. Consistent with their critical roles in regulatory biology, CRLs have been intimately linked to human disease pathogenesis and therapy: mutations in CRL subunits underlie the development of several human cancers and CRL4CRBN has been implicated as the target of the anti- myeloma agent Thalomid. CRL pathways are regulated by multiple mechanisms, including dynamic cycles of CRL assembly/disassembly and post-translational modification of the cullin subunit by the ubiquitin-like protein Nedd8. CSN is a Nedd8 isopeptidase that regulates CRLs by removing the Nedd8 modification. Removal of Nedd8 from a CRL results in its deactivation and renders its substrate receptor susceptible to disassembly by the exchange factor, Cand1. Although the isopeptidase activity of CSN inhibits CRL activity, genetic studies indicate that CSN is a positive regulator of CRLs, which suggests that periodic deactivation and disassembly of CRLs is required to sustain CRL activity. Recently, the Nedd8 conjugation pathway has emerged as a drug target for cancer chemotherapy and the active site subunit of Csn5 has been implicated in sustaining tumorigenicity in a Myc-driven cancer model. Together, these data point to dynamic cycling of CRLs as being critical for cancer cell proliferation. The over-arching hypotheses that animate this project are that CSN plays a critical role in regulating the activity of CRL enzymes and architecture of the CRL network, and that perturbation of CSN activity is toxic to cancer cells. We plan to explore these topics in three Specific Aims. In Aim 1 we will use quantitative multidimensional mass spectrometry to characterize the architecture and dynamics of the CRL proteome. In Aim 2, we will characterize a novel non-enzymatic CRL-inhibitory activity of CSN. In Aim 3, we will generate a CSN isopeptidase inhibitor to use as a research tool and to evaluate whether CSN is a promising target for chemotherapy of cancer.

描述（由申请人提供）：cullin-RING泛素连接酶（CRL）家族含有240种以上的泛素连接酶，调节广泛的细胞和生物过程。与其在调控生物学中的关键作用一致，CRL与人类疾病发病机制和治疗密切相关：CRL亚基中的突变是几种人类癌症发展的基础，并且CRL 4CRBN已被认为是抗骨髓瘤剂Thalomid的靶标。CRL途径受多种机制调节，包括CRL组装/拆卸的动态循环和泛素样蛋白Nedd 8对cullin亚基的翻译后修饰。CSN是一种Nedd 8异肽酶，通过去除Nedd 8修饰来调节CRL。从CRL中去除Nedd 8会导致其失活，并使其底物受体易于被交换因子Cand 1分解。虽然CSN的异肽酶活性抑制CRL活性，但遗传研究表明CSN是CRL的正调节剂，这表明需要定期灭活和拆卸CRL以维持CRL活性。最近，Nedd 8缀合途径已经成为癌症化疗的药物靶标，并且Csn 5的活性位点亚基已经涉及在Myc驱动的癌症模型中维持致瘤性。总之，这些数据表明CRL的动态循环对于癌细胞增殖至关重要。使该项目充满活力的过度假设是CSN在调节CRL酶的活性和CRL网络的结构中起关键作用，并且CSN活性的扰动对癌细胞是有毒的。我们计划在三个具体目标中探讨这些主题。目标1 我们将使用定量多维质谱来表征CRL蛋白质组的结构和动力学。在目标2中，我们将描述CSN的一种新的非酶促CRL抑制活性。在目标3中，我们将产生CSN异肽酶抑制剂，用作研究工具，并评估CSN是否是一个有前途的癌症化疗靶点。