Chemical Genetics of Transcriptional Regulation by CDKs in Human Cells
人类细胞中 CDK 转录调控的化学遗传学
基本信息
- 批准号:8630081
- 负责人:
- 金额:$ 32.21万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2014
- 资助国家:美国
- 起止时间:2014-02-15 至 2017-12-31
- 项目状态:已结题
- 来源:
- 关键词:Antineoplastic AgentsAntiviral AgentsArchaeal RNABindingCatalytic DomainCell Cycle ProgressionCell ProliferationCell divisionCellsChemicalsChromatinColon CarcinomaCyclin-Dependent KinasesDNA Polymerase IIDNA-Directed RNA PolymeraseDefectDevelopmentDrug TargetingElongation FactorEnsureEnzymesEukaryotaEvaluationFission YeastGene ExpressionGene Expression RegulationGenesGenetic ScreeningGenetic TranscriptionGenomeHumanHuman DevelopmentHuman EngineeringHuman IdentificationsIn VitroIndividualKineticsLabelMalignant NeoplasmsMessenger RNAModelingNormal tissue morphologyOrganismOrthologous GenePathway interactionsPatternPeptide Initiation FactorsPharmaceutical PreparationsPhasePhosphorylationPhosphotransferasesPlayPolymerasePositive Transcriptional Elongation Factor BProcessPropertyProteinsRNARNA Polymerase IIRNA ProcessingRNA chemical synthesisReadingRecruitment ActivityRegulationRoleSignal TransductionStagingTestingTimeTorpedoTranscriptTranscription ElongationTranscription InitiationTranscriptional RegulationVariantViralYeastsadenine analoganalogbasecancer cellchemical geneticschemotherapychromatin modificationchromatin proteincrosslinkcyclin T1cyclin-dependent kinase-activating kinasefeedinggenome-widehuman diseasein vivomutantnovelnovel strategiespreventpromoterpublic health relevancereconstitutionresearch studysmall moleculetranscription factor TFIIEtranscription factor TFIIHtranscription termination
项目摘要
Project Summary
The transcription cycle of RNA polymerase II (Pol II) depends on sequential functions of
distinct cyclin-dependent kinases (CDKs), but the mechanisms that order those functions
are still emerging. We have taken a chemical-genetic approach-replacement of wild-
type with analog-sensitive (AS) mutant CDKs-to reveal both "early" and "late" functions
of the CDK network in transcription. First, by selective inhibition of Cdk7-a component
of transcription initiation factor TFIIH-in human cells, we uncovered two unexpected,
and seemingly antagonistic functions. Cdk7 activity is required to recruit factors that
establish a promoter-proximal pause by Pol II, and to activate Cdk9, catalytic subunit of
positive transcription elongation factor b (P-TEFb), which releases the pause. Therefore
the CDK network appears to depend on incoherent feedforward to raise a transient
kinetic barrier to Pol II elongation, and thus create a temporal window to recruit mRNA-
processing and chromatin-modifying machinery. Consistent with a requirement for CDK-
directed pausing to ensure faithful RNA processing, inhibition of Cdk7 or Cdk9 leads to
defects in termination and 3'-end maturation of Pol II transcripts. Second, in a chemical-
genetic screen for Cdk9 substrates, we identified multiple proteins involved in RNA 5'-
end decapping and the "torpedo" pathway of transcription termination, which has
recently been suggested to influence pausing and divergent antisense transcription. We
will dissect the initiation-elongation transition of Pol II, and elucidate regulation of
transcription termination and polarity, to uncover novel modes of gene regulation by
CDKs. The specific aims are:
1. To identify functions and targets of CDKs at the initiation-elongation transition
2. To investigate possible regulation of the transcription termination pathway by P-
TEFb, uncovered in a chemical genetic screen for Cdk9 targets
3. To dissect functions of Cdk7 and Cdk9 by chemical genetics in human cells
Our studies reveal a CDK cascade at the core of the Pol II transcription cycle;
completion of our aims will illuminate how Cdk7 and Cdk9 collaborate to ensure
unidirectional transitions between phases of that cycle. By turning CDKs into chemical
switches that can be manipulated with customized small molecules, we will distinguish
their specific roles and substrates, and reveal new anti-cancer or anti-viral drug targets.
项目概要
RNA 聚合酶 II (Pol II) 的转录周期取决于以下功能的顺序:
不同的细胞周期蛋白依赖性激酶 (CDK),但控制这些功能的机制
仍在不断涌现。我们采取了化学遗传方法——替代野生——
具有模拟敏感 (AS) 突变体 CDK 的类型 - 揭示“早期”和“晚期”功能
CDK 网络在转录中的作用。一、通过选择性抑制Cdk7-a成分
在人类细胞中转录起始因子 TFIIH 的研究中,我们发现了两个意想不到的、
和看似对立的功能。 Cdk7 活性是招募因子所必需的
通过 Pol II 建立启动子近端暂停,并激活 Cdk9(催化亚基)
正转录延伸因子 b (P-TEFb),可释放暂停。所以
CDK 网络似乎依赖于不相干的前馈来引发瞬态
Pol II 延伸的动力学障碍,从而创建一个时间窗口来招募 mRNA-
加工和染色质修饰机械。符合 CDK 的要求-
定向暂停以确保忠实的 RNA 加工,抑制 Cdk7 或 Cdk9 会导致
Pol II 转录本的终止和 3' 端成熟缺陷。其次,在化学品中——
通过对 Cdk9 底物的遗传筛选,我们鉴定了多种参与 RNA 5'- 的蛋白质
末端脱帽和转录终止的“鱼雷”途径,
最近被认为影响反义转录的暂停和发散。我们
将剖析 Pol II 的起始-延伸转变,并阐明
转录终止和极性,通过以下方式揭示基因调控的新模式
CDK。具体目标是:
1. 确定CDK在起始-延伸转变过程中的功能和靶标
2. 研究P-转录终止途径的可能调控
TEFb,在 Cdk9 靶标的化学遗传筛选中发现
3. 通过化学遗传学剖析Cdk7和Cdk9在人体细胞中的功能
我们的研究揭示了 Pol II 转录周期核心的 CDK 级联;
我们目标的完成将阐明 Cdk7 和 Cdk9 如何合作以确保
该周期各阶段之间的单向转换。通过将 CDK 转化为化学物质
可以用定制的小分子操纵的开关,我们将区分
它们的特定作用和底物,并揭示新的抗癌或抗病毒药物靶点。
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)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
ROBERT P FISHER其他文献
ROBERT P FISHER的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('ROBERT P FISHER', 18)}}的其他基金
Cyclin-dependent kinase control of cell-division and transcription cycles
细胞分裂和转录周期的细胞周期蛋白依赖性激酶控制
- 批准号:
10559139 - 财政年份:2018
- 资助金额:
$ 32.21万 - 项目类别:
Cyclin-dependent kinase control of cell-division and transcription cycles
细胞分裂和转录周期的细胞周期蛋白依赖性激酶控制
- 批准号:
10370800 - 财政年份:2018
- 资助金额:
$ 32.21万 - 项目类别:
Cyclin-dependent kinase control of cell-division and transcription cycles
细胞分裂和转录周期的细胞周期蛋白依赖性激酶控制
- 批准号:
10378005 - 财政年份:2018
- 资助金额:
$ 32.21万 - 项目类别:
Cyclin-dependent kinase control of cell-division and transcription cycles
细胞分裂和转录周期的细胞周期蛋白依赖性激酶控制
- 批准号:
9903405 - 财政年份:2018
- 资助金额:
$ 32.21万 - 项目类别:
Chemical Genetics of Transcriptional Regulation by CDKs in Human Cells
人类细胞中 CDK 转录调控的化学遗传学
- 批准号:
8806563 - 财政年份:2014
- 资助金额:
$ 32.21万 - 项目类别:
Chemical Genetics of Transcriptional Regulation by CDKs in Human Cells
人类细胞中 CDK 转录调控的化学遗传学
- 批准号:
9198169 - 财政年份:2014
- 资助金额:
$ 32.21万 - 项目类别:
Chemical Genetic Analysis of the Human Cell Cycle
人类细胞周期的化学遗传分析
- 批准号:
8727082 - 财政年份:2013
- 资助金额:
$ 32.21万 - 项目类别:
Chemical Genetic Analysis of the Human Cell Cycle
人类细胞周期的化学遗传分析
- 批准号:
9128664 - 财政年份:2013
- 资助金额:
$ 32.21万 - 项目类别:
Chemical Genetic Analysis of the Human Cell Cycle
人类细胞周期的化学遗传分析
- 批准号:
8919920 - 财政年份:2013
- 资助金额:
$ 32.21万 - 项目类别:
Chemical Genetic Analysis of the Human Cell Cycle
人类细胞周期的化学遗传分析
- 批准号:
8479753 - 财政年份:2013
- 资助金额:
$ 32.21万 - 项目类别:
相似海外基金
Development of a new generation of antiviral agents that are effective against drug-resistant viruses and prevent serious illness and sequelae.
开发新一代抗病毒药物,可有效对抗耐药病毒并预防严重疾病和后遗症。
- 批准号:
23K18186 - 财政年份:2023
- 资助金额:
$ 32.21万 - 项目类别:
Grant-in-Aid for Challenging Research (Exploratory)
A versatile structure-based therapeutic platform for development of VHH-based antitoxin and antiviral agents
一个多功能的基于结构的治疗平台,用于开发基于 VHH 的抗毒素和抗病毒药物
- 批准号:
10560883 - 财政年份:2023
- 资助金额:
$ 32.21万 - 项目类别:
Genetically encoded bicyclic peptide libraries for the discoveryof novel antiviral agents
用于发现新型抗病毒药物的基因编码双环肽库
- 批准号:
10730692 - 财政年份:2021
- 资助金额:
$ 32.21万 - 项目类别:
Design and synthesis of nucleosides to develop antiviral agents and oligonucleotide therapeutics
设计和合成核苷以开发抗病毒药物和寡核苷酸疗法
- 批准号:
21K06459 - 财政年份:2021
- 资助金额:
$ 32.21万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Genetically encoded bicyclic peptide libraries for the discoveryof novel antiviral agents
用于发现新型抗病毒药物的基因编码双环肽库
- 批准号:
10189880 - 财政年份:2021
- 资助金额:
$ 32.21万 - 项目类别:
Computer-aided identification and synthesis of novel broad-spectrum antiviral agents
新型广谱抗病毒药物的计算机辅助鉴定和合成
- 批准号:
2404261 - 财政年份:2020
- 资助金额:
$ 32.21万 - 项目类别:
Studentship
Develop broad-spectrum antiviral agents against COVID-19 based on innate immune response to SARS-CoV-2 infection
基于对 SARS-CoV-2 感染的先天免疫反应,开发针对 COVID-19 的广谱抗病毒药物
- 批准号:
10222540 - 财政年份:2020
- 资助金额:
$ 32.21万 - 项目类别:
Develop broad-spectrum antiviral agents against COVID-19 based on innate immune response to SARS-CoV-2 infection
基于对 SARS-CoV-2 感染的先天免疫反应,开发针对 COVID-19 的广谱抗病毒药物
- 批准号:
10669717 - 财政年份:2020
- 资助金额:
$ 32.21万 - 项目类别:
Association between sedentary lifestyle and liver cancer development in hepatitis C patients treated with direct-acting antiviral agents
接受直接抗病毒药物治疗的丙型肝炎患者久坐的生活方式与肝癌发展之间的关系
- 批准号:
20K10713 - 财政年份:2020
- 资助金额:
$ 32.21万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Develop broad-spectrum antiviral agents against COVID-19 based on innate immune response to SARS-CoV-2 infection
基于对 SARS-CoV-2 感染的先天免疫反应,开发针对 COVID-19 的广谱抗病毒药物
- 批准号:
10174522 - 财政年份:2020
- 资助金额:
$ 32.21万 - 项目类别: