Structure, function, and regulation of the bacterial transcription cycle
Structure, function, and regulation of the bacterial transcription cycle
批准号:
10388954
负责人:
Seth A. Darst
金额:
$5.36万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
2016
资助国家:
美国
项目状态:
未结题
起止时间:
2016-05-01 至 2026-04-30
关键词:
AntibioticsBacteriaBacterial RNACatalytic DomainCombined Modality TherapyComplexCryoelectron MicroscopyCrystallizationDNADNA-Directed DNA PolymeraseDNA-Directed RNA PolymeraseDevelopmentDissociationElementsEnzymesFrequenciesGene ExpressionGenetic TranscriptionGoalsHoloenzymesLeadMacromolecular ComplexesProkaryotic CellsRNARegulationRifampicin resistanceRifampinRouteStructureTranscriptTuberculosisX-Ray Crystallographyantimicrobialinhibitor/antagonistinsightmacromolecular assemblymanpromoterresistant strainsmall moleculestructural biologythree dimensional structure
中文摘要
项目摘要
转录是基因表达的主要控制点,RNA聚合酶(RNAP),保守
从细菌到人类,都是转录的中心酶。我们的长期目标是了解
转录机制及其调控。确定RNAP和RNAP的三维结构
它与DNA、RNA和调节因子的复合体,是必不可少的一步。我们高度关注
鉴定了原核生物的RNAP。
阐明了转录周期的基本要素,即起始、延伸和终止
通过对原核生物的研究。对整个的结构和功能有详细的了解
转录循环是解释基因表达的基本控制和靶向的关键
RNAP加小分子抗生素。这一理解的进展受困于
可视化瞬变中间体,这些中间体是稳定状态之间关键转变的基础
转录循环,以及可视化复杂的大分子组装涉及的困难
管理,结构问题,其中X射线结晶学有严重的限制。
而稳定的RNAP状态在转录周期周围(RNAP催化核心,RNAP
全酶、RNAP全酶开放启动子复合体、RNAP延伸复合体)相对
人们很好地描述和理解了稳定状态之间的转变,但对这些转变的理解却很少。
主要过渡包括:
全酶启动子dnaè开放启动子复合体(启动)
开放启动子复合体è延伸复合体(启动子逃逸,σ解离)
延长复合体è核心RNAPDNA完成rna转录本(终止)
这些转变中的每一个都具有不稳定的、瞬时的中间产物,这些中间产物非常
对结构生物学的挑战。
在转录周期的每个阶段,RNAP的功能都受到与外源基因的相互作用的调节
监管因素。含有外源基因的转录复合体的组装和结晶
监管机构也给结构生物学带来了挑战。
由于最近的进展,低温电子显微镜(CRYO-EM)现在提供了一种结构和
这些中间体和大型组件的机械表征。我们将使用低温电子
显微镜,结合X射线结晶学和其他方法来利用这一点
机会,并提供细菌转录周期的完整特征。
英文摘要
Project Summary
Transcription is the major control point of gene expression and RNA polymerase (RNAP), conserved
from bacteria to man, is the central enzyme of transcription. Our long term goal is to understand the
mechanism of transcription and its regulation. Determining three-dimensional structures of RNAP and
its complexes with DNA, RNA, and regulatory factors, is an essential step. We focus on highly
characterized prokaryotic RNAPs.
The basic elements of the transcription cycle, initiation, elongation, and termination, were elucidated
through study of prokaryotes. A detailed structural and functional understanding of the entire
transcription cycle is essential to explain the fundamental control of gene expression and to target
RNAP with small-molecule antibiotics. Advances in this understanding are stuck on the difficulty of
visualizing transient intermediates that underlie the key transitions between stable states of the
transcription cycle, and the difficulty of visualizing complex macromolecular assemblies involved in
regulation, structural problems where X-ray crystallography has severe limitations.
While the stable RNAP states around the transcription cycle (RNAP catalytic core, RNAP
holoenzyme, RNAP holoenzyme open promoter complex, RNAP elongation complex) are relatively
well characterized and understood, the transitions between the stable states are poorly understood.
Major transitions include:
Holoenzyme + promoter DNA è open promoter complex (initiation)
Open promoter complex è elongation complex (promoter escape, σ dissociation)
Elongation complex è core RNAP + DNA + completed RNA transcript (termination)
Each of these transitions are characterized by unstable, transient intermediates that are extremely
challenging for structural biology.
At every stage of the transcription cycle, RNAP function is modulated by interactions with extrinsic
regulatory factors. Assembling and crystallizing transcription complexes containing extrinsic
regulators also presents challenges for structural biology.
Due to recent advances, cryo-electron microscopy (cryo-EM) now offers a route to structural and
mechanistic characterization of these intermediates and large assemblies. We will use cryo-electron
microscopy, in combination with X-ray crystallography and other approaches, to exploit this
opportunity and provide a complete characterization of the bacterial transcription cycle.
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会议论文
Structure, function, and regulation of the bacterial transcription cycle
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批准号:10607993
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项目类别:
-
资助金额:$83.56万
-
财政年份:2016
-
负责人:Seth A. Darst
-
依托单位:
Structure, function, and regulation of the bacterial transcription cycle
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批准号:10394344
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项目类别:
-
资助金额:$83.56万
-
财政年份:2016
-
负责人:Seth A. Darst
-
依托单位:
Structure, function, and regulation of the bacterial transcription cycle
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批准号:9921406
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项目类别:
-
资助金额:$81.12万
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财政年份:2016
-
负责人:Seth A. Darst
-
依托单位:
Structure, function, and regulation of the bacterial transcription cycle
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批准号:9071516
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项目类别:
-
资助金额:$79.1万
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财政年份:2016
-
负责人:Seth A. Darst
-
依托单位:
Structure, function, and regulation of the bacterial transcription cycle
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批准号:9271202
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项目类别:
-
资助金额:$81.12万
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财政年份:2016
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负责人:Seth A. Darst
-
依托单位:
Structural studies of RNA polymerase regulation by RNA
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批准号:8238020
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项目类别:
-
资助金额:$34.02万
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财政年份:2012
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负责人:Seth A. Darst
-
依托单位:
Structural studies of RNA polymerase regulation by RNA
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批准号:8431355
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项目类别:
-
资助金额:$31.83万
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财政年份:2012
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负责人:Seth A. Darst
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依托单位:
Structural studies of RNA polymerase regulation by RNA
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批准号:8794441
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项目类别:
-
资助金额:$32.98万
-
财政年份:2012
-
负责人:Seth A. Darst
-
依托单位:
Structural studies of RNA polymerase regulation by RNA
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批准号:8608542
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项目类别:
-
资助金额:$32.98万
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财政年份:2012
-
负责人:Seth A. Darst
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依托单位:
?/ANTI-? COMPLEXES: STAPHYLOCOCCAL AUREUS PHAGE G1 ORF67
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批准号:8169306
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项目类别:
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资助金额:$0.2万
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财政年份:2010
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负责人:Seth A. Darst
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依托单位:
STRUCTURAL STUDIES OF BACTERIAL SIGNALLING: SPORULATION CONTROL
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批准号:8169240
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项目类别:
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资助金额:$0.2万
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财政年份:2010
-
负责人:Seth A. Darst
-
依托单位:
A GENERIC METHOD TO STUDY BACTERIOPHAGE/HOST INTERACTIONS
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批准号:8169128
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项目类别:
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资助金额:$0.12万
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财政年份:2010
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负责人:Seth A. Darst
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依托单位:
STRUCTURAL BASIS FOR MICROTUBULE CROSSLINKING BY THE MAP65 PROTEIN FAMILY
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批准号:8169310
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项目类别:
-
资助金额:$0.2万
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财政年份:2010
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负责人:Seth A. Darst
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依托单位:
Bacterial RNAP sigma factor structure and function
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批准号:8055634
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项目类别:
-
资助金额:$2.26万
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财政年份:2010
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负责人:Seth A. Darst
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依托单位:
CRYSTAL STRUCTURES OF THE GLYCOPEPTIDE SULFOTRANSFERASE TEG12
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批准号:8169309
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项目类别:
-
资助金额:$0.2万
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财政年份:2010
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负责人:Seth A. Darst
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依托单位:
STRUCTURE OF A PAUSED TRANSCRIPTION ELONGATION COMPLEX
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批准号:8169308
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项目类别:
-
资助金额:$0.2万
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财政年份:2010
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负责人:Seth A. Darst
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依托单位:
BACTERIAL RNA POLYMERASE ? FACTOR INTERACTIONS WITH THE PROMOTER -10 ELEMENT
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批准号:8169305
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项目类别:
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资助金额:$0.2万
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财政年份:2010
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负责人:Seth A. Darst
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依托单位:
DETERMINE THE STRUCTURAL BASIS FOR ?N INTERACTIONS WITH ITS PROMOTER DNA
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批准号:8169307
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项目类别:
-
资助金额:$0.2万
-
财政年份:2010
-
负责人:Seth A. Darst
-
依托单位:
A GENERIC METHOD TO STUDY BACTERIOPHAGE/HOST INTERACTIONS
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批准号:7954085
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项目类别:
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资助金额:$0.59万
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财政年份:2009
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负责人:Seth A. Darst
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依托单位:
STRUCTURAL STUDIES OF BACTERIAL SIGNALLING: SPORULATION CONTROL
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批准号:7955130
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项目类别:
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资助金额:$2.5万
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财政年份:2009
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负责人:Seth A. Darst
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依托单位:
国内基金
海外基金
Segmented Filamentous Bacteria激活宿主免疫系统抑制其拮抗菌 Enterobacteriaceae维持菌群平衡及其机制研究
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批准号:81971557
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项目类别:面上项目
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资助金额:65.0万元
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批准年份:2019
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负责人:毛开睿
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依托单位:
电缆细菌(Cable bacteria)对水体沉积物有机污染的响应与调控机制
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批准号:51678163
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资助金额:64.0万元
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批准年份:2016
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负责人:许玫英
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依托单位: