Mechanism of Transcript Elongation in Chromatin by RNA Polymerase II
RNA 聚合酶 II 染色质转录本延伸机制
基本信息
- 批准号:8456094
- 负责人:
- 金额:$ 1.83万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:1999
- 资助国家:美国
- 起止时间:1999-01-01 至 2013-06-30
- 项目状态:已结题
- 来源:
- 关键词:AddressAffectBiochemicalBiological AssayChromatinChromatin StructureComplexComputer AnalysisDNADNA RepairDNA biosynthesisDataDevelopmentDissociationElongation FactorEnvironmentEnzymesEukaryotic CellFOS geneFluorescence Resonance Energy TransferFluorescent ProbesGene Expression RegulationGenesGenetic TranscriptionGoalsHIVHealthHistone H2AHistone H4HistonesHumanIn VitroMalignant NeoplasmsMetabolismMolecularMolecular ChaperonesMolecular GeneticsMono-SNucleosomesOutcomePlayPoint MutationPolymerasePositioning AttributePrincipal InvestigatorProcessProteinsProto-OncogenesRNA Polymerase IIRecoveryRegulationRegulator GenesResearchRoleStructural ModelsStructureSystemTechniquesTestingTimeTranscriptTranscription InitiationTranscription Processc-myc Geneschromatin modificationchromatin remodelingclinically significanthistone modificationhuman diseaseimprovedin vivoleukemianormal agingparticleprogramspromoterprotein protein interactionresearch studysingle moleculetime usetumorigenesis
项目摘要
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
描述(申请人提供):RNA聚合酶II(pol II)转录的重要过程发生在真核细胞的染色质环境中;事实上,中度转录的基因保留了核小体结构。最近的研究表明,染色质结构为体外转录pol II提供了一个强大的屏障,并且在中等活性基因的转录延长过程中,DNA-组蛋白相互作用仅部分和短暂地被破坏。此外,延长pol II复合物是基因调控过程中的主要靶标之一。核小体屏障和与组蛋白和pol II相互作用的蛋白质因子参与这种调节。这些研究提出了以下问题:(1)真核生物聚合酶如何克服核小体屏障?(2)在pol II通过染色质的过程中,组蛋白的恢复机制是什么?(3)与染色质和靶酶相互作用的因子如何改变pol II通过染色质的进展速率?本项目的长期研究目标是了解染色质中pol II对转录延长的调控机制。具体的问题将在一个高度纯化的体外转录系统中解决。我们将使用生物化学,荧光,分子遗传学和单分子技术的组合分析同质和明确的单核和多核染色质模板的转录。我们的实验将集中在分析真核细胞pol II。我们的初步研究表明,通过染色质的转录伴随着瞬时DNA从组蛋白上解螺旋和组蛋白八聚体的解折叠。这些中间体的结构和它们之间的相互转化率在很大程度上决定了通过染色质转录过程的结果。因此,具体目标是:1。通过pol II染色质转录过程中DNA-组蛋白相互作用的变化以及中间体之间的相互转化率提供分子描述。2.鉴定转录依赖的组蛋白八聚体解折叠过程中蛋白质-蛋白质相互作用的扰动,以及参与该过程的因素。PHS 398/2590(Rev.06/09)
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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VASILY M STUDITSKY其他文献
VASILY M STUDITSKY的其他文献
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{{ truncateString('VASILY M STUDITSKY', 18)}}的其他基金
Mechanisms of PARP-1 interaction with chromatin
PARP-1 与染色质相互作用的机制
- 批准号:
9382154 - 财政年份:2017
- 资助金额:
$ 1.83万 - 项目类别:
Mechanism of Transcript Elongation in Chromatin by RNA Polymerase II
RNA 聚合酶 II 染色质转录本延伸机制
- 批准号:
8831693 - 财政年份:1999
- 资助金额:
$ 1.83万 - 项目类别:
Mechanism of Transcript Elongation in Chromatin by RNA Polymerase II
RNA 聚合酶 II 染色质转录本延伸机制
- 批准号:
8692320 - 财政年份:1999
- 资助金额:
$ 1.83万 - 项目类别:
Mechanism of Transcript Elongation in Chromatin by RNA Polymerase II
RNA 聚合酶 II 染色质转录本延伸机制
- 批准号:
8297105 - 财政年份:1999
- 资助金额:
$ 1.83万 - 项目类别:
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