Structural insight into the role of CHD4 PHD fingers.

对 CHD4 PHD 手指作用的结构洞察。

• 批准号: 7675771
• 负责人: Catherine Anne Musselman
• 金额: $ 5.01万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7675771
• 关键词: ATP phosphohydrolase; Affinity; Arrhythmia; Autoantigens; Autoimmune Diseases; Binding; Binding Proteins; Binding Sites; Biochemical; Biological Assay; Chemicals; Chromatin; Complex; DNA-Binding Proteins; Data; Deacetylase; Development; Diagnostic; Disease Marker; Etiology; Fingers; Fluorescence Microscopy; Fluorescence Spectroscopy; Green Fluorescent Proteins; Heart Arrest; Heart failure; Histone H3; Histones; Human; In Situ; Individual; Knowledge; Length; Ligands; Link; Measures; Molecular; Molecular Biology; Mutagenesis; Myocardial Infarction; Myositis; N-terminal; NMR Spectroscopy; Nature; Nucleosomes; PHD Finger; Peptides; Proteins; Public Health; Regulation; Repression; Research; Resolution; Risk; Role; Solutions; Specificity; Structure; Tail; Testing; Transcriptional Regulation; Western Blotting; X-Ray Crystallography; base; chromatin immunoprecipitation; helicase; in vivo; insight; mutant; novel; prognostic; protein crosslink; three dimensional structure

DESCRIPTION (provided by applicant): Human CHD4 (chromodomain helicase DMA-binding protein 4) is an autoantigen of myositis, an autoimmune disease associated with an increased risk of conjestive heart failure, arrhythmia, myocardial infarction and cardiac arrest. CHD4 is an ATPase and a major subunit of the NuRD (nucleosome remodeling and deacetylase) repression complex, which is involved in transcriptional regulation and development. The role of CHD4 in this complex, however, remains unclear. Our preliminary data suggest that two PHD fingers of CHD4 recognize histone H3, with PHD2 specifically recognizing H3K9me3, a novel interaction. We hypothesize that the tandem PHD fingers of CHD4 bind to the N-terminal tail of histone H3, tethering the CHD4/NuRD complex to chromatin. We seek to elucidate the molecular basis and the functional significance of the CHD4 PHD-H3 interaction. The specific aims are: 1) To establish the molecular mechanism of histone recognition by the CHD4 PHD1 and PHD2 fingers, and 2) To elucidate the role of the tandem PHD (PHD1.2) domain of CHD4 in histone recognition. A set of structural, biochemical and molecular biology approaches, including NMR spectroscopy, X-ray crystallography, mutagenesis and in vivo chromatin immunoprecipitation and fluorescence microscopy will be used to determine the three dimensional structures of the CHD4 PHD fingers and to probe their interaction with histone H3 peptides. Fluorescence spectroscopy and NMR will be used to measure the binding affinities and to determine whether the two PHD fingers bind cooperatively. PUBLIC HEALTH RELEANCE: The results generated in this study will provide a fundamental knowledge about functioning of the CHD4/NuRD complex and will thus aid in determining the etiology of myositis and may help to identify new prognostic and diagnostic markers of this disease.

描述（由申请人提供）：人类CHD4（染色体蛋白酶DMA结合蛋白4）是一种肌炎的自身抗原，这是一种自身免疫性疾病，伴随着伴有心律失常，心律失常，心肌梗塞和心脏骤停的风险增加。 CHD4是ATPase和NURD（核小体重塑和脱乙酰基酶）抑制复合物的主要亚基，它参与了转录调控和发育。然而，CHD4在该复合物中的作用尚不清楚。我们的初步数据表明，CHD4的两个PHD手指识别组蛋白H3，PHD2专门识别H3K9me3，这是一种新型的相互作用。我们假设CHD4的串联PHD手指与组蛋白H3的N末端尾巴结合，将CHD4/NURD复合物链接到染色质。我们寻求阐明CHD4 PHD-H3相互作用的分子基础和功能意义。具体目的是：1）建立CHD4 PHD1和PHD2手指的组蛋白识别的分子机制，以及2）阐明CHD4在组蛋白识别中的串联PHD（PHD1.2）结构域的作用。一组结构，生化和分子生物学方法，包括NMR光谱，X射线晶体学，诱变和体内染色质免疫沉淀和荧光显微镜，可用于确定CHD4 PHD手指的三维结构，并探测其与组酮H3肽的相互作用。荧光光谱和NMR将用于测量结合亲和力，并确定两个PHD手指是否合作结合。公共卫生恢复：本研究产生的结果将提供有关CHD4/NURD Complex的功能的基本知识，因此将有助于确定肌炎的病因，并可能有助于确定该疾病的新预后和诊断标记。