RNA 聚合酶 II 染色质转录本延伸机制-猫眼课题宝

RNA 聚合酶 II 染色质转录本延伸机制

批准号：

8831693

负责人：

VASILY M STUDITSKY

金额：

$36.59万

依托单位：

RESEARCH INST OF FOX CHASE CAN CTR

依托单位国家：

美国

项目类别：

财政年份：

1999

资助国家：

美国

项目状态：

已结题

起止时间：

1999-01-01 至 2017-01-31

来源：

https://reporter.nih.gov/project-details/8831693

关键词：

Address Affect Biochemical Biological Assay Chromatin Chromatin Structure Complex Computer Analysis DNA DNA Repair DNA biosynthesis Data Development Dissociation Elongation Factor Environment Enzymes Eukaryotic Cell FOS gene Fluorescence Resonance Energy Transfer Fluorescent Probes Gene Expression Regulation Genes Genetic Transcription Goals HIV Health Histone H2A Histone H4 Histones Human In Vitro Malignant Neoplasms Metabolism Molecular Molecular Chaperones Molecular Genetics Mono-S Nucleosomes Outcome Play Point Mutation Polymerase Positioning Attribute Principal Investigator Process Proteins Proto-Oncogenes RNA Polymerase II Recovery Regulation Regulator Genes Research Role Structural Models Structure System Techniques Testing Time Transcript Transcription Initiation Transcription Process c-myc Genes chromatin modification chromatin remodeling clinically significant histone modification human disease improved in vivo leukemia normal aging particle programs promoter protein protein interaction research study single molecule time use tumorigenesis

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DESCRIPTION (provided by applicant): The vital process of transcription by RNA polymerase II (pol II) occurs in chromatin environment in eukaryotic cells; in fact, moderately transcribed genes retain nucleosomal structure. Recent studies suggest that chromatin structure presents a strong barrier for transcribing pol II in vitro, and that DNA-histone interactions are only partialy and transiently disrupted during transcript elongation on moderately active genes. Furthermore, elongating pol II complex is one of the major targets during gene regulation. The nucleosomal barrier and protein factors interacting with histones and pol II participate in this regulation. These studies raise the following questions: (1) How do eukaryotic polymerases overcome the nucleosome barrier? (2) What are the mechanisms of histone recovery during progression of pol II through chromatin? (3) How do the factors interacting with chromatin and with the target enzymes change the rate of pol II progression through chromatin? The long-term research goal of this proposal is to understand the mechanism and the regulation of transcript elongation by pol II in chromatin. The specific questions will be addressed in a highly purified transcription system in vitro. We will analyze transcription of homogeneous and well-defined mono- and polynucleosomal chromatin templates using a combination of biochemical, fluorescent, molecular genetic and single-molecule techniques. Our experiments will be focused on analysis of eukaryotic pol II. Our preliminary studies have shown that transcription through chromatin is accompanied by transient DNA uncoiling from histones and unfolding of histone octamer. The structures of these intermediates and the rates of interconversion between them largely determine the outcome of the process of transcription through chromatin. Accordingly, the specific aims are: 1. To provide molecular description of the changes in DNA-histone interactions during transcription through chromatin by pol II and the rates of interconversion between the intermediates. 2. To identify protein-protein interactions perturbed during transcription-dependent unfolding of the histone octamer, and factors involved in this process. PHS 398/2590 (Rev. 06/09) Page Continuation Format Page

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