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Single Molecule Studies of Transcription Complexes

转录复合物的单分子研究

基本信息

批准号：
7558821
负责人：
Steven Chu
金额：
$ 20.03万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/7558821
关键词：
Address Binding Biochemical Color Complement Complex DNA DNA Binding DNA Sequence DNA-Directed RNA Polymerase Detection Distal Distant Drosophila genus Elements Enhancers Eukaryota Eukaryotic Cell Event Fluorescence Microscopy Fluorescent Dyes Gene Expression General Transcription Factors Genes Genetic Transcription Glass Human In Vitro Incubated Indium Individual Label Magnetism Measurement Measures Mediating Methodology Methods Modeling Molecular Monitor Names Numbers Polymerase Principal Investigator Process Proteins RNA Polymerase II Rate Recruitment Activity Regulatory Element Relative (related person)Signal Transduction Site Standards of Weights and Measures Surface System Techniques Testing Time Transcription Coactivator Transcription Initiation Transcriptional Activation base cancer therapy cofactor genetic regulatory protein melting polypeptide programs promoter research study single molecule

项目摘要

Transcription of protein-encoding genes in eukaryotes is a highly regulated process that is initiated when a myriad of transcriptional activators recognize specific DNA elements proximal and distal to core promoters. These DNA bound activators then direct the dynamic assembly of RNA polymerase II, general transcription factors, and cofactor complexes on promoter DNA to initiate transcription. As the exact sequence of events needed to initiate transcription is poorly understood, a mechanistic understanding of how transcription activators regulate and direct transcription complex formation from their proximal and distant recognition sites is crucial to understanding how genes expression is regulated. Toward a better understanding of the mechanism of RNAPII mediated transcription we will develop and apply single molecule techniques to (i) measure the rates and the sequence by which the GTFs and RNAPII bind to promoter DNA, (ii) determine how the rates and the sequence by which the GTFs and RNAPII bind to promoter DNA are modulated by transcriptional activators and co- activators, and (iii) determine how activation occurs by distal enhancers. Information on dynamics of transcription initiation obtained in these single-molecule studies will complement the structural information obtained by the proposed biochemical, EM, and x-ray crystallographic studies of this PPG. Mechanistic understanding of transcription complex formation will provide new potential protein targets for anticancer therapies.

在真核生物中，蛋白质编码基因的转录是一个高度调控的过程