Single-molecule visualization of transcription regulation mechanisms

转录调控机制的单分子可视化

• 批准号: 7931231
• 负责人: JEFF GELLES
• 金额: $ 12.28万
• 依托单位: BRANDEIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7931231
• 关键词: Affect; Bacteria; Binding; Binding Sites; Biochemical; Biology; Cell Differentiation process; Cell physiology; Chemicals; Communicable Diseases; Complex; DNA; DNA Binding; DNA-Directed RNA Polymerase; Data; Development; Disease; Dyes; Enhancers; Equilibrium; Escherichia coli; Eukaryota; Fluorescence; Fluorescence Microscopy; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genomics; Human; Human Biology; Imagery; In Vitro; Individual; Kinetics; Knowledge; Label; Laboratories; Malignant Neoplasms; Measures; Mediating; Molecular; Molecular Machines; Monitor; Normal Cell; Nucleotides; Organism; Pathway interactions; Polymerase; Population; Population Study; Prokaryotic Cells; Property; Proteins; Public Health; Reaction; Regulation; Regulator Genes; Repression; Research; Research Personnel; Site; Stimulus; System; Technology; Testing; Time; Transcript; Transcription Elongation; Transcription Initiation; Transcriptional Activation; Transcriptional Regulation; Virulence; base; brain cell; combat; genetic regulatory protein; improved; insight; instrumentation; interest; novel; novel strategies; pathogen; programs; promoter; prototype; research study; response; single molecule; transcription factor

DESCRIPTION (provided by applicant): A central concern of the present post-genomic era of biology is understanding the chemical and physical mechanisms by which gene expression is regulated. Appropriate activation and repression of particular genes is necessary for maintaining normal cell function and is required for executing the programs of cell differentiation that are essential to the development of multicellular organisms. Collectively, gene regulatory systems are the "brain" of the cell that allow it to respond appropriately to environmental stimuli. Many cancers and other diseases result from deranged gene regulation. We here propose an entirely new approach to studying the molecular mechanisms of gene regulation in vitro. Instead of studying populations of molecules, we will directly visualize the regulatory machinery attached to an isolated single DNA molecule, following the progression of the machinery through its different states in real time while simultaneously observing the extent of transcriptional activation. Such direct visualization is made possible by novel multi-wavelength single-molecule fluorescence instrumentation newly developed our laboratory. This approach will allow us for the first time to elucidate regulation mechanisms by directly analyzing the dynamics of individual molecular interactions in complete regulatory complexes, instead of relying on inferences founded on data from piecemeal studies on individual proteins and their equilibrium interactions with DNA or with RNA polymerase. We will apply this technology to three different systems involved in regulation of transcription initiation and elongation in Escherichia coli. Each system was chosen because it is a prototype for a common mechanism of transcription regulation that functions analogously in both prokaryotes and eukaryotes. The proposed research will elucidate basic mechanisms of transcription regulation. In the long term this will improve public health by improving our understanding of human biology. In addition, the proposed research will help define the molecular basis for regulatory switches that affect virulence and environmental dissemination of human pathogens. This basic knowledge is expected to aid in the scientific research aimed at development of agents to combat infectious disease.

描述（由申请人提供）：当前后基因组时代生物学的一个中心问题是理解基因表达被调控的化学和物理机制。特定基因的适当激活和抑制是维持正常细胞功能所必需的，也是执行细胞分化程序所必需的，而细胞分化是多细胞生物发育所必需的。总的来说，基因调控系统是细胞的“大脑”，它允许细胞对环境刺激做出适当的反应。许多癌症和其他疾病都是由于基因调控失常造成的。