Nucleosome Reorganizing/Remodeling Complexes

核小体重组/重塑复合物

• 批准号: 7580908
• 负责人: CHRISTOPHER P. HILL
• 金额: $ 25.15万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2010-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7580908
• 关键词: ATP phosphohydrolase; Actins; Affinity; Binding; Binding Proteins; Biochemical; Biochemical Genetics; Biochemistry; Biological; Biological Assay; Biological Process; Biological Testing; Bromodomain; Chromatin; Cocrystallography; Collaborations; Columbidae; Complex; Crystallography; DNA; DNA Polymerase II; DNA biosynthesis; Data; Foundations; Genes; Genetic Screening; Genetic Transcription; Goals; Histones; Homologous Gene; Human; In Vitro; Individual; Knowledge; Length; Link; Lysine; Maintenance; Maps; Mediating; Modeling; Molecular; Nucleosomes; Phase; Physiology; Polymerase; Principal Investigator; Procedures; Process; Productivity; Protein Subunits; Proteins; RNA; RNA Processing; Replication Origin; Repression; Research Design; Resolution; Risk; Role; Structure; Sum; Tail; Taiwan; Transcription Elongation; Transcription Initiation; Transcription Process; Translating; Yeasts; base; biophysical chemistry; degradosome; design; high reward; in vivo; milligram; nuclease; pathogen; programs; promoter; protein complex; success; yeast genetics

Nucleosome structure has profound implications for DNA-templated processes and modulation of this structure is of critical importance for replication and transcription. Nucleosome structure must be loosened (reorganized) to facilitate polymerase passage and normal structure must be re-established in the wake of the elongating polymerase. Also, nucleosomes must be translated along DNA (remodeled) to allow access to promoter sequences and replication origins. Our goal is to understand the molecular basis for these activities, and our proposal is organized into three specific Aims, each of which focuses on a different protein or complex: Spt16-Pob3 (Aim 1) is a reorganizing complex that restructures nucleosomes to facilitate transcription initiation and passage of elongating RNA Pol II. It also performs an important (presumably equivalent) role in DNA replication. Spt6 (Aim 2) performs a related role to Spt16-Pob3 by promoting reformation of normal nucleosome structure following passage of RNA Pol II. It also provides a link between the machineries for RNA transcription and processing. RSC (Aim 3) is an ATP driven remodeling complex that functions in the expression of numerous genes by mediating nucleosomal translocation. Our approach is highly collaborative and broad. Because our goal is to arrive at a detailed molecular understanding we will determine relevant structures by x-ray crystallography (Hill, PI). Because we aim to understand processes in a biological context we are exploiting the strengths of yeast genetics (Formosa, co- I; and Cairns, collaborator). The link between structural and in vivo studies is provided by biochemical analysis, which is ongoing in all three labs. Further strength is added by our collaboration with experts in biophysical chemistry (David Myszka) and pathogen physiology and biochemistry (Simon Dove). We have made considerable progress toward the specific aims. In each case important structures have been determined and biochemical and genetic assays developed. Preliminary data indicate that considerable productivity will ensue and also lay the foundation for longer-term goals that carry more risk but also offer potential for high reward.

核小体结构对DNA-模拟过程具有深远的影响和对此的调节 结构对于复制和转录至关重要。核小体结构必须松动 （重组）以促进聚合酶通过和正常结构，必须在 细长聚合酶。此外，必须沿DNA（重塑）翻译核小体以允许进入 启动子序列和复制起源。我们的目标是了解这些分子基础 活动，我们的提议分为三个特定目标，每个目标都集中在不同的蛋白质上 或复合物：SPT16-POB3（AIM 1）是一种重组的复合物，可重组核小体以促进 伸长RNA Pol II的转录启动和通过。它还执行重要（大概是 等效）在DNA复制中的作用。 SPT6（AIM 2）通过促进与SPT16-POB3发挥相关作用 RNA POL II通过后正常核小体结构的改革。它还提供了 RNA转录和处理的机器。 RSC（AIM 3）是ATP驱动的重塑复合物 通过介导核小体易位，该作用在众多基因的表达中。 我们的方法是高度协作和广泛的。因为我们的目标是到达详细的分子 了解我们将通过X射线晶体学（Hill，PI）确定相关结构。因为我们的目标是 在生物学环境中了解过程，我们正在利用酵母遗传学的优势（福尔摩萨，共同 我;和Cairns，合作者）。结构和体内研究之间的联系由生化提供 分析，这在所有三个实验室中都在进行。我们与专家的合作增加了进一步的优势 生物物理化学（David Myszka）和病原体生理和生物化学（Simon Dove）。 我们已经朝着具体目标取得了长足的进步。在每种情况下，重要结构都有 已确定并开发了生化和遗传测定。初步数据表明 将会发生相当大的生产率，并为长期目标奠定基础，这些目标具有更大的风险，但 还提供了高奖励的潜力。