Chromatin Regulation by Deacetylation and SUMO-Targeted Ubiquitin Ligation

通过脱乙酰化和 SUMO 靶向泛素连接调节染色质

• 批准号: 7883402
• 负责人: LORRAINE PILLUS
• 金额: $ 31.16万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7883402
• 关键词: Acetylation; Acute Promyelocytic Leukemia; Affect; Age; Aging; Architecture; Binding; Biochemical; Biochemical Genetics; Biological; Biological Assay; Biology; Cancer Control; Candidate Disease Gene; Cell Cycle Regulation; Cells; Chromatin; Chromosomal Rearrangement; Chromosome Structures; Complex; DNA; DNA Damage; DNA biosynthesis; Deacetylase; Deacetylation; Defect; Development; Disease; Enzymes; Epigenetic Process; Family; Genetic; Genetic Recombination; Genetic Transcription; Genetic screening method; Genome Stability; Genomics; Goals; Growth; Health; Histones; Human; Human Activities; Human Development; In Vitro; Lead; Ligation; Link; Malignant Neoplasms; Mediator of activation protein; Metabolic Diseases; Metabolism; Microscopic; Modification; Molecular; Molecular Genetics; Muscle; Nuclear; Pattern; Post-Translational Protein Processing; Process; Proteins; Proteomics; Recombinant DNA; Regulation; Research; Role; Sampling; Site; Structural Protein; Structure; Substrate Specificity; Syndrome; Testing; Therapeutic; Therapeutic Agents; Therapeutic Intervention; Transcriptional Activation; Ubiquitin; Yeasts; cell growth; cell type; chromatin immunoprecipitation; detection of nutrient; dietary restriction; genetic analysis; genome wide association study; healthy aging; in vivo; insight; mutant; novel; protein function; public health relevance; recombinational repair; repaired; research study; response

DESCRIPTION (provided by applicant): Chromosomal functions are regulated by dynamic modifications of their structural proteins. Acetylation and deacetylation are well-established modifications that affect how histones and other chromosomal proteins influence transcription, recombination, replication and repair of damage. A distinct, newly defined activity is SUMO-targeted ubiquitin ligation (STUbL), catalyzed by proteins previously identified for their roles in genome stability and in response to DNA. The goals of the research are to define the mechanisms by which chromatin deacetylation and STUbL together yield optimal transcriptional silencing, genome stability and growth regulation. The proposed research builds on the lab's recent discovery that the Sir2 deacetylase is physically and functionally linked to a STUbL activity catalyzed by the Slx5-Slx8 complex. The project will be accomplished through three aims. In the first aim, genetic and biochemical studies will test the hypothesis that a second predicted STUbL protein functions in parallel to SLX5-SLX8 to promote optimal structure and function through STUbL activity. Transcriptional silencing defects of mutants will be characterized through molecular and genetic approaches, including genetic analysis and chromatin immunoprecipitation (ChIP). In the second aim, the genomic and subnuclear localization of STUbL components will be evaluated. Genomic and cell biological experiments will test if STUbLs occupy silent chromatin and define a distinct genomic binding pattern and subnuclear compartment(s). The third aim will define chromatin-specific substrates of STUbL activity through biochemical approaches. A combination of candidate substrate and proteomic analyses will be used. Potential substrates will be independently validated through additional biochemical and molecular genetic approaches. It will be determined if Sir2 deacetylase activity influences STUbL activity or substrate specificity. Together, the results from these three aims will establish the key mechanisms, substrates, and genomic targets of STUbL that are critical for chromatin function. PUBLIC HEALTH RELEVANCE: The STUbL enzymes that are the focus of this study affect critical processes such as cell growth, aging, metabolism, and response to DNA damage. In human cells, a STUbL was recently shown to be responsible for the therapeutic benefits in treatments of acute promyelocytic leukemia, yet many unanswered questions remain about this newly identified family of enzymes. Understanding their cellular roles, substrates and regulation will ultimately provide insight into basic processes that define human development and disease, and may be particularly relevant to mechanisms of healthy aging and cancer.

描述（由申请人提供）：染色体功能通过其结构蛋白的动态修饰来调节。乙酰化和脱乙酰化是公认的修饰，影响组蛋白和其他染色体蛋白如何影响转录、重组、复制和损伤修复。一种独特的、新定义的活性是 SUMO 靶向泛素连接 (STUbL)，由先前确定的在基因组稳定性和 DNA 响应中发挥作用的蛋白质催化。该研究的目标是确定染色质脱乙酰化和 STUbL 共同产生最佳转录沉默、基因组稳定性和生长调节的机制。拟议的研究建立在实验室最近发现的基础上，即 Sir2 脱乙酰酶在物理和功能上与 Slx5-Slx8 复合物催化的 STUbL 活性相关。该项目将通过三个目标来实现。在第一个目标中，遗传和生化研究将检验以下假设：第二个预测的 STUbL 蛋白与 SLX5-SLX8 并行发挥作用，通过 STUbL 活性促进最佳结构和功能。突变体的转录沉默缺陷将通过分子和遗传学方法进行表征，包括遗传分析和染色质免疫沉淀（ChIP）。第二个目标是评估 STUbL 成分的基因组和亚核定位。基因组和细胞生物学实验将测试 STUbL 是否占据沉默染色质并定义独特的基因组结合模式和亚核区室。第三个目标是通过生化方法定义 STUbL 活性的染色质特异性底物。将使用候选底物和蛋白质组分析的组合。潜在的底物将通过额外的生化和分子遗传学方法进行独立验证。将确定 Sir2 脱乙酰酶活性是否影响 STUbL 活性或底物特异性。这三个目标的结果将共同确定对染色质功能至关重要的 STUbL 的关键机制、底物和基因组靶标。 公共健康相关性：本研究重点关注的 STUbL 酶影响细胞生长、衰老、代谢和 DNA 损伤反应等关键过程。在人类细胞中，STUbL 最近被证明对治疗急性早幼粒细胞白血病具有治疗效果，但关于这个新发现的酶家族仍有许多未解答的问题。了解它们的细胞作用、底物和调节最终将有助于深入了解定义人类发育和疾病的基本过程，并且可能与健康衰老和癌症的机制特别相关。