权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Mechanisms of reversible DUB oxidation in genome stability pathways

基因组稳定性途径中可逆 DUB 氧化的机制

基本信息

批准号：
9145183
负责人：
Tony Tung Huang
金额：
$ 38.14万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-30 至 2020-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9145183
关键词：
Active Sites Affect Allosteric Regulation Biochemical Biological Assay Biological Models Cancer Etiology Caspase Cell physiology Cells Chemicals Chronic Cysteine DNA Repair Data Defect Deubiquitinating Enzyme Deubiquitination Development Disease Equilibrium Family Fanconi&apos s Anemia Genome Stability Genomic Instability Health Human Human Genome In Vitro Link Maintenance Malignant Neoplasms Mammalian Cell Mass Spectrum Analysis Mediating Modification Monoubiquitination Normal Cell Oxidation-Reduction Oxidative Stress Pathway interactions Pharmacotherapy Play Proteins Reactive Oxygen Species Regulation Resistance Role Signal Transduction Site Ubiquitin Ubiquitination Work base cancer therapy cell growth environmental agent in vivo mutant neoplastic cell novel oxidation response sensor signal processing therapeutic target tumorigenesis ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): The deregulation of genome stability pathways caused by environmental agents is a major culprit of tumorigenesis in human cells; protein ubiquitination, a reversible posttranslational signaling process, is essential for maintaining this regulation. The ubiquitination status of a target protein is achieved via a delicate balance between two opposing forces: ubiquitin E3 ligases and deubiquitinating enzymes (DUBs). It has been postulated that the majority of proteins in a cell are regulated and modified by ubiquitin at some point; however, due to the multitude of DUBs present in cells, many of the modifications only exist transiently and are difficult to capture. There are nearly one hundred DUBs encoded in the human genome, many of which have uncharacterized function(s) and regulation. DUBs play an essential role in the maintenance of genomic stability, and their aberrant expression has been linked to tumorigenesis. The regulation of DUBs in mammalian cells and how this affects genome stability pathways remain unclear. DUBs constitute a large family of cysteine (Cys) proteases yet basic information on how the enzymatic activity of DUBs is modulated by oxidative stress caused by either environmental agents or intracellular signals is lacking. We recently discovered that bursts of reactive oxygen species (ROS) reversibly inactivate specific DUBs through the oxidation of the catalytic Cys residue. Importantly, the DUB USP1, a key regulator of the Fanconi Anemia (FA) genomic stability pathway and PCNA-mediated translesion synthesis (TLS), was reversibly inactivated upon oxidative stress. We hypothesize that DUBs of the cysteine protease family act as ROS sensors in human cells and that ROS-mediated DUB inactivation is a critical mechanism for fine-tuning genome stability pathways in both health and disease. Herein, we propose to dissect the different mechanisms of DUB regulation using USP1 as a model system, and then extend the analysis to other Cys- and metallo-based DUBs. We will elucidate the mechanisms and functions of DUB oxidation by using a set of unique biochemical and mass spectrometry-based assays to capture and characterize the reversibly oxidized form(s) of DUBs. We will investigate the role of DUB oxidation in DNA repair and cell growth pathways by generating DUB mutants that remain active, yet resistant to reversible oxidation. Finally, we will employ a unique approach to collect and study novel ubiquitinated species that are regulated by oxidative stress. Together, our studies will reveal core principles of deubiquitination and genome stability regulation, and will likely provide guidance for the development of novel classes of therapeutics that target DUBs.