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: 9188804
• 负责人: SETH M COHEN
• 金额: $ 32.77万
• 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/9188804
• 关键词: Active Sites; Affinity Chromatography; Antineoplastic Agents; Architecture; Behavior; Biochemical; Biological; Biological Assay; Biology; COPS5 gene; 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; Genetic study; Guanine Nucleotide Exchange Factors; HDAC1 gene; Human; IL27RA gene; 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; Periodicity; 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; biophysical analysis; cancer cell; cancer therapy; carbonate dehydratase; experimental study; genetic approach; human disease; inhibitor/antagonist; isopeptidase; lenalidomide; novel; overexpression; preclinical study; preference; public health relevance; receptor; repository; response; scaffold; small molecule libraries; targeted treatment; 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(申请人提供)：剔除环泛素连接酶(CRL)家族包含240多种泛素连接酶，调节广泛的细胞和生物过程。CRL在调节生物学中的关键作用与人类疾病的发病机制和治疗密切相关：CRL亚单位的突变是几种人类癌症发生的基础，CRL4CRBN已被认为是抗骨髓瘤药物Thalomid的靶点。CRL途径受多种机制的调控，包括CRL组装/拆解的动态循环和泛素样蛋白Nedd8对cullin亚基的翻译后修饰。CSN是一种Nedd8异肽酶，通过去除Nedd8修饰来调节CRL。从CRL中去除Nedd8会导致其失活，并使其底物受体容易被交换因子Cand1拆解。尽管CSN的异肽酶活性抑制CRL的活性，但遗传学研究表明CSN是CRL的正调节因子，这表明CRL需要周期性的失活和解体才能维持CRL活性。最近，Nedd8结合途径已成为癌症化疗的药物靶点，Csn5的活性部位亚单位在Myc驱动的癌症模型中参与维持肿瘤的致瘤性。总而言之，这些数据表明CRL的动态循环对癌细胞增殖至关重要。该项目的主要假设是CSN在调节CRL酶的活性和CRL网络的结构方面发挥关键作用，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下探讨这些主题。在目标1中 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，我们将表征CSN的一种新的非酶CRL抑制活性。在目标3中，我们将生产一种CSN异肽酶抑制剂作为研究工具，并评估CSN是否为癌症化疗的有前景的靶点。

项目成果

Epidithiodiketopiperazines Inhibit Protein Degradation by Targeting Proteasome Deubiquitinase Rpn11.

桥二硫二酮哌嗪通过靶向蛋白酶体去泛素酶 Rpn11 抑制蛋白质降解。

• DOI: 10.1016/j.chembiol.2018.07.012
• 发表时间: 2018-11-15
• 期刊: Cell chemical biology
• 影响因子: 8.6
• 作者: Li J;Zhang Y;Da Silva Sil Dos Santos B;Wang F;Ma Y;Perez C;Yang Y;Peng J;Cohen SM;Chou TF;Hilton ST;Deshaies RJ
• 通讯作者: Deshaies RJ

Targeting Metalloenzymes for Therapeutic Intervention.

• DOI: 10.1021/acs.chemrev.8b00201
• 发表时间: 2019-01-23
• 期刊: Chemical reviews
• 影响因子: 62.1
• 作者: Chen AY;Adamek RN;Dick BL;Credille CV;Morrison CN;Cohen SM
• 通讯作者: Cohen SM

Alcohol-abuse drug disulfiram targets cancer via p97 segregase adaptor NPL4.

• DOI: 10.1038/nature25016
• 发表时间: 2017-12-14
• 期刊: Nature
• 影响因子: 64.8
• 作者: Skrott Z;Mistrik M;Andersen KK;Friis S;Majera D;Gursky J;Ozdian T;Bartkova J;Turi Z;Moudry P;Kraus M;Michalova M;Vaclavkova J;Dzubak P;Vrobel I;Pouckova P;Sedlacek J;Miklovicova A;Kutt A;Li J;Mattova J;Driessen C;Dou QP;Olsen J;Hajduch M;Cvek B;Deshaies RJ;Bartek J
• 通讯作者: Bartek J

Structure-Activity Relationships in Metal-Binding Pharmacophores for Influenza Endonuclease.

• DOI: 10.1021/acs.jmedchem.8b01363
• 发表时间: 2018-11-21
• 期刊: Journal of medicinal chemistry
• 影响因子: 7.3
• 作者: Credille CV;Dick BL;Morrison CN;Stokes RW;Adamek RN;Wu NC;Wilson IA;Cohen SM
• 通讯作者: Cohen SM

A Bioinorganic Approach to Fragment-Based Drug Discovery Targeting Metalloenzymes.

• DOI: 10.1021/acs.accounts.7b00242
• 发表时间: 2017-08-15
• 期刊: Accounts of chemical research
• 影响因子: 18.3
• 作者: Cohen SM