Epigenetic regulation by PWWP domain proteins

PWWP 结构域蛋白的表观遗传调控

• 批准号: 7895337
• 负责人: Songtao Jia
• 金额: $ 22.3万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-10 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7895337
• 关键词: Acetylation; Adaptor Signaling Protein; Affect; Affinity Chromatography; Amino Acids; Antibodies; Binding; Biological Assay; Biological Process; Cell physiology; Chromatin; Chromatin Structure; Complex; Coupled; Defect; Disease; Enzymes; Epigenetic Process; Epitopes; Family; Fission Yeast; Gene Targeting; Genes; Goals; Histone Acetylation; Histones; In Vitro; Lead; Link; Mass Spectrum Analysis; Mediating; Methylation; Methyltransferase; Modification; Mutagenesis; Mutation; PWWP Domain; Pattern; Peptides; Phenotype; Phosphorylation; Play; Process; Proteins; Recruitment Activity; Regulation; Role; Specificity; Substrate Specificity; Western Blotting; base; chromatin immunoprecipitation; design; developmental disease; genome-wide; histone acetyltransferase; histone methyltransferase; histone modification; human disease; immunodeficiency-centromeric instability-facial anomalies syndrome; in vivo; insight; member; novel therapeutic intervention; protein complex; protein function

DESCRIPTION (provided by applicant): Covalent modification of histones, such as acetylation, methylation, phosphorylation, and ubiquitylation, are essential regulators of chromatin structure and function. Defects in the regulation of these modifications have causal roles in numerous developmental disorders and diseases. However, the mechanisms that regulate histone-modifying activities are not well understood. In addition, apart from a few well-known examples, it is not clear how the cellular machinery interprets these modifications. The PWWP domain is similar to Chromo, Tudor, and MBT domains, which are well known for their ability to recognize methylated histones to regulate diverse cellular processes. However, the function of the PWWP domain is still a mystery. Our preliminary studies demonstrated that PWWP domain proteins directly interact with histones and form complexes with histone-modifying activities. I therefore hypothesize that PWWP domain proteins might recognize modified chromatin components to regulate associated enzymes. I will characterize the functions of PWWP domain proteins in fission yeast to gain mechanistic insights into their roles in regulating chromatin structure and function. The specific aims are designed to (a) determine the histone modification that interacts with each PWWP domain protein with pull-down assays and chromatin immunoprecipitation (ChIP) analysis; (b) characterize PWWP domain protein complexes identified by affinity purification of epitope-tagged PWWP domain proteins to elucidate the contribution of each protein to the function of these complexes; (c) analyze the cellular functions of PWWP domain proteins in chromatin based processes, with the goal of directly linking phenotypes to specific histone modifications. Mutations in many PWWP domain-containing proteins are intimately linked to human diseases. For example, a single amino acid alteration in the PWWP domain of Dnmt3b is responsible for ICF (immunodeficiency, centromeric instability, and facial anomalies) syndrome. A better understanding of the function of PWWP domain proteins may lead to novel therapeutic approaches to treat these diseases.

描述（由申请人提供）：组蛋白的共价修饰，例如乙酰化，甲基化，磷酸化和泛素化，是染色质结构和功能的必不可少的调节剂。这些修饰的调节缺陷在许多发育障碍和疾病中具有因果关系。但是，调节组蛋白改良活性的机制尚不清楚。此外，除了一些著名的示例外，尚不清楚蜂窝机械如何解释这些修饰。 PWWP结构域类似于Chromo，Tudor和MBT结构域，它们以识别甲基化组蛋白以调节各种细胞过程的能力而闻名。但是，PWWP域的功能仍然是一个谜。我们的初步研究表明，PWWP结构域蛋白直接与组蛋白相互作用，并与组蛋白修饰活性形成复合物。因此，我假设PWWP结构域蛋白可能识别改良的染色质成分以调节相关酶。我将表征裂变酵母中pwwp结构蛋白的功能，以使其在调节染色质结构和功能中的作用中获得机械见解。具体目的旨在（a）确定与每个PWWP结构蛋白相互作用的组蛋白修饰，并通过下拉分析和染色质免疫沉淀（CHIP）分析； （b）表征通过对表位标记的PWWP结构蛋白的亲和力纯化来鉴定的PWWP结构蛋白蛋白复合物，以阐明每种蛋白质对这些复合物功能的贡献； （c）分析基于染色质过程中PWWP结构蛋白的细胞功能，其目的是将表型直接连接到特定的组蛋白修饰。许多含PWWP结构域蛋白质的突变与人类疾病密切相关。例如，DNMT3B的pWWP结构域中的单个氨基酸改变是ICF（免疫缺陷，丝粒不稳定和面部异常）综合征。更好地了解pwwp结构蛋白的功能可能会导致新的治疗方法来治疗这些疾病。