Mechanisms of Chromatin Transcriptional Regulation

染色质转录调控机制

• 批准号: 9276390
• 负责人: Michael Guy Poirier
• 金额: $ 5万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-02-01 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9276390
• 关键词: Acetylation; Alzheimer' s Disease; Binding; Binding Sites; Biochemical; Bladder Neoplasm; Chemicals; Chromatin; Chromatin Remodeling Factor; Colorectal Neoplasms; Complex; DNA; Defect; Dissociation; Drug Targeting; Drug resistance; Epigenetic Process; Excision; Fluorescence; Fluorescence Spectroscopy; Foundations; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Genome Stability; Goals; Health; Histone H3.3; Histones; Human Genome; Kinetics; Location; Lysine; Malignant Neoplasms; Mammary Neoplasms; Measurement; Measures; Mediating; Methodology; Molecular; Molecular Chaperones; N-terminal; Nature; Nerve Degeneration; Nucleosomes; Oncogenes; Positioning Attribute; Post-Translational Protein Processing; Protein Chemistry; Regulation; Site; Structure; Tail; Transcription Elongation; Transcription Initiation; Transcriptional Regulation; Tyrosine Phosphorylation; Variant; cancer therapy; chromatin modification; chromatin remodeling; deep sequencing; design; epigenetic drug; histone modification; novel anticancer drug; ovarian neoplasm; promoter; single molecule; tool; transcription factor; tumorigenesis

DESCRIPTION (provided by applicant): Alterations in gene expression cause numerous cancers including ovarian, bladder, breast and colorectal tumors, as well as neurodegenerative conditions, such as Alzheimer's disease. The dynamic organization of the human genome into chromatin regulates transcription initiation and elongation, while defects in chromatin modifications, assembly, disassembly and remodeling results in the misregulation of many oncogenes. This has led to chromatin modifying complexes becoming one of the main chemotherapeutic drug targets. Numerous studies have identified and correlated the components involved in chromatin transcriptional regulation (CTR), which include histone variants, histone post-translational modifications (PTMs), histone chaperone assembly factors and chromatin remodeling factors. Remarkably, genetic, biochemical, structural and deep sequencing studies have not fully revealed the mechanisms of CTR. We hypothesize that nucleosome unwrapping, histone PTMs, histone variants, histone chaperones and chromatin remodelers function together to significantly influence TF occupancy and H2A-H2B heterodimer exchange to regulate transcription. We have developed precise chemical tools and single molecule measures capable of determining the mechanisms and functions of CTR. Using these methodologies, we will determine the mechanisms by which histone variants, PTMs, chaperones and chromatin remodeling complexes function together to cooperatively (additively and multiplicatively), anti-cooperatively and redundantly regulate transcription. We propose in the renewal application the following Specific Aims: (1) Determine the mechanisms by which nucleosomes influence transcription factor dynamics; (2) Determine the transcriptional regulatory mechanisms of H2A.Z, H3.3 and nucleosome entry-exit PTMs. (3) Determine the transcriptional regulator mechanisms behind H2A-H2B heterodimer exchange and removal. These studies will provide a foundation for understanding the mechanisms and functional interactions behind chromatin transcriptional regulation and the misregulation of numerous oncogenes.

描述(申请人提供)：基因表达的改变导致多种癌症，包括卵巢、膀胱癌、乳腺癌和结直肠癌，以及神经退行性疾病，如阿尔茨海默病。人类基因组到染色质的动态组织调节转录的起始和延伸，而染色质修饰、组装、拆解和重塑的缺陷导致许多癌基因的错误调节。这使得染色质修饰络合物成为主要的化疗药物靶点之一。大量研究已经确定并关联了参与染色质转录调节(CTR)的组分，包括组蛋白变体、组蛋白翻译后修饰(PTM)、组蛋白伴侣组装因子和染色质重塑因子。值得注意的是，遗传、生化、结构和深度测序研究尚未完全揭示CTR的机制。我们假设，核小体解包装、组蛋白PTM、组蛋白变体、组蛋白伴侣蛋白和染色质重构体共同发挥作用，显著影响TF的占有率和H_2A-H_2B异源二聚体交换来调节转录。我们已经开发出能够确定CTR机制和功能的精确化学工具和单分子测量方法。使用这些方法，我们将确定组蛋白变体、PTM、伴侣和染色质重塑复合体协同(相加和乘法)、反协同和冗余调控转录的机制。我们在更新应用中提出了以下具体目标：(1)确定核小体影响转录因子动态的机制；(2)确定H2A.Z、H3.3和核小体进出PTM的转录调控机制。(3)确定H_2A-H_2B异源二聚体交换和去除背后的转录调控机制。这些研究将为理解染色质转录调控和多种癌基因的错误调控背后的机制和功能相互作用提供基础。

项目成果

Histone H3 phosphorylation near the nucleosome dyad alters chromatin structure.

• DOI: 10.1093/nar/gku150
• 发表时间: 2014-04
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: North JA;Šimon M;Ferdinand MB;Shoffner MA;Picking JW;Howard CJ;Mooney AM;van Noort J;Poirier MG;Ottesen JJ
• 通讯作者: Ottesen JJ

Sequence-dependent thymine dimer formation and photoreversal rates in double-stranded DNA.

• DOI: 10.1039/c3pp50078k
• 发表时间: 2013-08
• 期刊: Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology
• 作者: Law YK;Forties RA;Liu X;Poirier MG;Kohler B
• 通讯作者: Kohler B

Structural basis for high-affinity binding of LEDGF PWWP to mononucleosomes.

• DOI: 10.1093/nar/gkt074
• 发表时间: 2013-04-01
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Eidahl JO;Crowe BL;North JA;McKee CJ;Shkriabai N;Feng L;Plumb M;Graham RL;Gorelick RJ;Hess S;Poirier MG;Foster MP;Kvaratskhelia M
• 通讯作者: Kvaratskhelia M

Chemical and biological tools for the preparation of modified histone proteins.

• DOI: 10.1007/128_2015_629
• 发表时间: 2015
• 期刊: Topics in current chemistry
• 影响因子: 8.6
• 作者: Howard CJ;Yu RR;Gardner ML;Shimko JC;Ottesen JJ
• 通讯作者: Ottesen JJ

Preparation of fully synthetic histone H3 reveals that acetyl-lysine 56 facilitates protein binding within nucleosomes.

• DOI: 10.1016/j.jmb.2011.01.003
• 发表时间: 2011-04-29
• 期刊: Journal of molecular biology
• 影响因子: 5.6
• 作者: Shimko JC;North JA;Bruns AN;Poirier MG;Ottesen JJ
• 通讯作者: Ottesen JJ