Physical Chemistry of Nucleic Acids

核酸物理化学

基本信息

批准号：
8300854
负责人：
CARLOS Jose BUSTAMANTE
金额：
$ 35.52万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
1983
资助国家：
美国
起止时间：
1983-07-01 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8300854
关键词：
Affect Base Pairing Behavior Biological Assay Cells Charge Complex Coupled DNA DNA-Directed RNA Polymerase Development Diffusion Elongation Factor Enzymes Escherichia coli Event Future Gene Expression Gene Expression Regulation Generations Genes Genetic Transcription Health Histone Acetylation Histones Image Imaging Techniques Ionic Strengths Kinetics Laws Lead Mechanics Modeling Motor Movement NURF Nucleic Acids Nucleosome Core Particle Nucleosomes Nucleotides Pathology Pathway interactions Physical Chemistry Physical condensation Polymerase Post-Translational Protein Processing Process RNA RNA Polymerase II Reaction Resolution Rotation Time Torque Torsion Transcription Elongation Yeasts analog chromatin remodeling congenital heart disorder density dimer experience histone modification in vivo insight laser tweezer mutant nucleotide analog particle research study single molecule transcription factor S-II tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): Transcription constitutes the central point of control for gene expression in the cell. The eukaryotic Pol II enzyme must transcribe in the presence of the physical barriers present in nucleosomal DNA and both this enzyme and its prokaryotic counterpart must transcribe torsionally constrained DNA segments that require the enzymes to translocate against increasing torques. Moreover, during transcription elongation these two highly structurally homologous enzymes perform a mechanochemical cycle in which a nucleotide condensation reaction is coupled to the mechanical tasks of movement, force, and torque generation. In this application we propose to use a combination of imaging and single molecule manipulation approaches to: a) Investigate the dynamics of transcription through nucleosomal DNA by yeast Pol II. b) Determine the effect of elongation factors on nucleosomal transcription. c) Investigate the effect of histone modification on the dynamics of nucleosomal transcription. d) Investigate the effect of remodeling factors on the dynamics of Pol II through nucleosomal DNA. We also propose to continue our previous studies of the prokaryotic E. coli RNA polmerase enzyme to investigate: a) Its generation of torque during transcription of torsionally constrained DNA, and b) The details of its mechanochemical cycle at high spatial resolution. PUBLIC HEALTH RELEVANCE: Transcription constitutes the central point of control for regulation of gene expression. Disruption of this process is associated with congenital heart disease, oncogenesis, and a large variety of developmental pathologies. The insights that our experiments provide on transcription will support the first steps in the development of future therapies.

描述（由申请人提供）：转录构成了细胞中基因表达的控制点。真核生物pol II酶必须在核小体DNA中存在的物理屏障的情况下转录，该酶及其原核对应物必须转录扭转的约束DNA片段，这些DNA片段要求酶需要这些酶反对增加扭矩。此外，在转录伸长期间，这两种高度结构同源的酶执行了机械化学循环，其中核苷酸凝结反应与运动，力和扭矩产生的机械任务耦合。在此应用中，我们建议将成像和单分子操纵方法的组合使用： b）确定伸长因子对核小体转录的影响。 c）研究组蛋白修饰对核小体转录动力学的影响。 d）研究重塑因子对POL II通过核小体DNA的动力学的影响。我们还建议继续我们先前对原核生物大肠杆菌RNA polmerase酶的研究，以研究：a）其在扭转约束的DNA转录过程中的产生，b）其机械化学周期在高空间分辨率下的细节。公共卫生相关性：转录构成了调节基因表达的控制点。该过程的破坏与先天性心脏病，肿瘤发生和多种发育病理有关。我们的实验对转录提供的见解将支持未来疗法开发的第一步。