Selective inhibition of CDK7 reveals high-confidence targets and new models for TFIIH function in transcription.
Selective inhibition of CDK7 reveals high-confidence targets and new models for TFIIH function in transcription.
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DOI：
10.1101/gad.341545.120
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发表时间：
2020-11-01
影响因子：
10.5
通讯作者：
Taatjes DJ
中科院分区：
文献类型：
--
作者：
Rimel JK;Poss ZC;Erickson B;Maas ZL;Ebmeier CC;Johnson JL;Decker TM;Yaron TM;Bradley MJ;Hamman KB;Hu S;Malojcic G;Marineau JJ;White PW;Brault M;Tao L;DeRoy P;Clavette C;Nayak S;Damon LJ;Kaltheuner IH;Bunch H;Cantley LC;Geyer M;Iwasa J;Dowell RD;Bentley DL;Old WM;Taatjes DJ
在这项研究中，Rimel等人着手研究CDK 7在转录中的作用。使用基于SILAC的磷酸蛋白质组学与转录组学和生物化学测定，作者鉴定了高置信度的CDK 7底物，这是剪接中对CDK 7活性的令人惊讶的广泛需求，以及涉及其与TFIIH相关的CDK 7激酶调控的意想不到的方面。CDK 7与10-亚基TFIIH复合物结合，并通过磷酸化RNA聚合酶II（RNAPII）的C-末端结构域（CTD）来调节转录。很少有其他CDK 7底物是已知的。在这里，使用共价抑制剂SY-351和定量磷酸蛋白质组学，我们确定了CDK 7激酶底物在人类细胞。在数百个高置信度目标中，绝大多数是CDK 7所特有的（即，与其他转录相关激酶不同），具有提示新细胞功能的子集。转录相关因子是主要的CDK 7底物，包括SF 3B 1、U2 AF 2和其他剪接组分。因此，广泛和多样的剪接缺陷，如选择性外显子包含和内含子保留，其特征在于CDK 7抑制细胞。结合生物化学测定，我们确定CDK 7直接激活其他转录相关激酶CDK 9，CDK 12和CDK 13，在转录中发挥“主调节”作用。我们进一步证明TFIIH将CDK 7激酶功能限制于RNAPII CTD，而其他底物（例如，SPT 5和SF 3B 1）被三亚基CDK激活激酶（CAK; CCNH、MAT 1和CDK 7）磷酸化。这些结果提示了CDK 7在转录中功能的新模型，并暗示CAK从TFIIH解离对于激酶活化是必需的。这种简单的调控策略确保CDK 7激活在空间和时间上与转录相关，并可能适用于其他转录相关激酶。
In this study, Rimel et al. set out to investigate the roles of CDK7 in transcription. Using SILAC-based phosphoproteomics with transcriptomics and biochemical assays, the authors identified high-confidence CDK7 substrates, a surprisingly widespread requirement for CDK7 activity in splicing, and unexpected aspects of CDK7 kinase regulation that involve its association with TFIIH. CDK7 associates with the 10-subunit TFIIH complex and regulates transcription by phosphorylating the C-terminal domain (CTD) of RNA polymerase II (RNAPII). Few additional CDK7 substrates are known. Here, using the covalent inhibitor SY-351 and quantitative phosphoproteomics, we identified CDK7 kinase substrates in human cells. Among hundreds of high-confidence targets, the vast majority are unique to CDK7 (i.e., distinct from other transcription-associated kinases), with a subset that suggest novel cellular functions. Transcription-associated factors were predominant CDK7 substrates, including SF3B1, U2AF2, and other splicing components. Accordingly, widespread and diverse splicing defects, such as alternative exon inclusion and intron retention, were characterized in CDK7-inhibited cells. Combined with biochemical assays, we establish that CDK7 directly activates other transcription-associated kinases CDK9, CDK12, and CDK13, invoking a “master regulator” role in transcription. We further demonstrate that TFIIH restricts CDK7 kinase function to the RNAPII CTD, whereas other substrates (e.g., SPT5 and SF3B1) are phosphorylated by the three-subunit CDK-activating kinase (CAK; CCNH, MAT1, and CDK7). These results suggest new models for CDK7 function in transcription and implicate CAK dissociation from TFIIH as essential for kinase activation. This straightforward regulatory strategy ensures CDK7 activation is spatially and temporally linked to transcription, and may apply toward other transcription-associated kinases.
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DOI：
10.1038/nrm.2017.63
发表时间：
2017-10
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
Herzel L;Ottoz DSM;Alpert T;Neugebauer KM
通讯作者：
Neugebauer KM
人 TFIIH 激酶 CDK7 调节转录相关的染色质修饰。
影响因子：
8.8
作者：
Ebmeier CC;Erickson B;Allen BL;Allen MA;Kim H;Fong N;Jacobsen JR;Liang K;Shilatifard A;Dowell RD;Old WM;Bentley DL;Taatjes DJ
通讯作者：
Taatjes DJ
DOI：
10.1073/pnas.0903642106
发表时间：
2009-08-25
影响因子：
11.1
作者：
Hong, Sun Woo;Hong, Seong Min;Lee, Dong-ki
通讯作者：
Lee, Dong-ki
影响因子：
8.8
作者：
Darman, Rachel B.;Seiler, Michael;Buonamici, Silvia
通讯作者：
Buonamici, Silvia
影响因子：
16.6
作者：
Compe, Emmanuel;Genes, Carlos M.;Egly, Jean-Marc
通讯作者：
Egly, Jean-Marc