TFIIH consists of two subcomplexes: the core complex, comprising ATP-dependent helicases, and the CAK complex, which harbours the kinase activity of CDK7 [ 8 ]. Decreased CAK activity creates a feedback loop, which turns off TFIIH activity. Transcription factor IIH (TFIIH) is a multiprotein complex involved in both eukaryotic transcription and DNA repair, revealing a tight connection between these two processes. Surprisingly, inhibition of TFIIH kinase activity only partially affected both Pol II density and Ser-2 phosphorylation level. Figure 1: TFIIH(CAK) CTD kinase activity binds to the VP16 activation domain. These results suggest that the TFIIH kinase activity is essential for This mechanism ensures the fidelity of chromosome transmission. the kinase activity, are critical for global gene transcription. Here we show that human TFIIH has DNA-dependent ATPase activity and we characterize the stimulatory effect of TFIIE on both the ATPase and kinase activities. For an activated promoter, targets of repression are TFIID and TFIIH, while for a basal promoter, TFIIH is the major target for mitotic inactivation of transcription. CAK also plays a role in DNA damage response. The kinase activity of TFIIH displayed stronger substrate preference for Cdk4 than did CAK. TFIIH consists of a complex of 10 subunits, 3 of which including the CDK7 kinase form a subcomplex called CAK ( 28 ). TFIIH, a 10-subunit complex with many resident enzymatic activities, is essential for transcription by RNA polymerase II (Pol II) ( 1 5 ). Kin28, the TFIIH kinase subunit, is important for promoter escape not elongation Kin28 depletion dramatically increases Mediator occupancy at the core promoter Stronger defects in promoter escape are linked to stronger transcriptional effects TFIIH phosphorylation of the CTD causes Mediator dissociation and promoter escape Summary The substrate specificities ofTFIIH and two forms of the Cdk7containing kinase complex are compared, and the relationship between transcription activity and the TFIIHdependent phosphorylation of the carboxy terminal domain of the largest subunit of PolII (CTD) is studied. CDK7 is the serine/threonine kinase of the basal transcription factor TFIIH. Background The general transcription factor TFIIH plays important roles in initiation and the transition to elongation steps of transcription by RNA polymerase II (PolII). The transcription/DNA repair factor TFIIH consists of nine subunits, several exhibiting known functions: helicase/ATPase, kinase activity and DNA binding. Transcription factor II Human is an important protein complex, having roles in transcription of various protein-coding genes and DNA nucleotide excision repair pathways. Download : Download high-res image (444KB) TFIIH is a eukaryotic complex composed of two subcomplexes, the CAK (Cdk Activating Kinase) and the core-TFIIH. Both roles are TAF7 inhibited the phosphorylation of the Pol II CTD by the CAK complex, demonstrating that TAF7 acts directly on CAK ( Fig. TFIIH, a large protein complex consisting of 10 subunits, is functionally divided into two subcomplexes ( Table 1) [99]: the seven-subunit core, which contains three disease-related gene products, XPB ( ERCC3 ), XPD ( ERCC2 ), and TTDA ( GTF2H5) [100102]; and the CDK-activating kinase (CAK) module, composed of CDK7, cyclin H, and MAT1 [103,104]. Inhibition of CAK prevents cell cycle from progressing. 1995). Moreover, TFIIH-phosphorylated CTD stimulated SETD1A/B activity toward nucleosomes, revealing a mechanistic basis for CDK7 regulation of H3K4me3 spreading. We observed a strong negative correlation between the gene-expression ratio and transcription rate. [1] TFIIH along with Pol II and several other multisubunit complexes assembles into a preinitiation complex These and other observations to be discussed below strongly suggest that the loss of Kin28 kinase, rather than the TFIIH complex, results in a defect in promoter escape, such that Pol II spends relatively more time at or near the PIC in the Kin28-depleted strain than in the control strain. A kinase activity specific for CTD is a component of the general transcription factor TFIIH. Repression of TFIIH Transcriptional Activity and TFIIH-Associated cdk7 Kinase Activity at Mitosis TFIIE stimulates the TFIIH-dependent kinase activity that phosphorylates the carboxy-terminal domain of the largest subunit of RNA polymerase II 6, and possesses a The general transcription factor TFIIH plays important roles in initiation and the transition to Requirement for TFIIH kinase activity in transcription by RNA polymerase II Sasha Akoulitchev, Tomi P. Mkel, Robert A. Weinberg & Danny Reinberg Nature 377 , HeLa nuclear extract was fractionated on equivalent GST-VP16 or control GST affinity columns. These sections were identical in different eukaryotic organisms from yeast to humans, suggesting that these regions may be important for tuning or controlling the activity 1995). TFIIH is also part of the DNA repair machinery and phosphorylates both cdc2 and cdk2 (Shiekhattar et al. 1 A ). The discrepancy between mRNA level and Pol II density is attributed to the defective 5-capping, which results in the destabilization of mRNAs. We also show that transcription requires CTD kinase activity provided by the CDK7 subunit of TFIIH17-19. It is shown that the CTD, but not its phosphoryla-tion, is required for initiation of Here the substrate specicities During the transcription, TFIIH enters the RNA TFIIH along with Pol II and several enzyme activities of TFIIH contribute to transcrip-tion has remained elusive. for the genes transcribed by rna polymerase iii (pol iii) (such as 5s rrna and trna genes), previous studies have documented that the activity of the general class iii transcription factor tfiiib is greatly diminished in extracts from synchronized mitotic cells (49) or by the conversion of an interphase xenopusegg extract to the mitotic state by TFIIH consists of nine subunits, and one of them, Cdk7, possesses kinase activity. Its activities include DNA-dependent ATPase helicase, C-terminal domain kinase, catalyzation of promoter escape, and participation in NER. 1994; Shiekhattar et al. The columns The kinase subunit (Kin28 in S. cerevisiae or Cdk7 in humans) marks the transcriptionally active polymerase by hyperphosphorylating the C-terminal domain (CTD) of its largest subunit ( Lee and Young, 2000, Liu et al., 2013, Roeder, 1996 ). TAF7 inhibited the CTD kinase activities of TFIIH ( Fig. Three subunits of TFIIH, cdk7, cyclin H and MAT1, form a ternary complex, cdk-activating kinase (CAK), found either on its own or as part of TFIIH. We [1] The activity of CAK associated with TFIIH decreases when DNA is damaged by UV irradiation. (A) Transcription of the CMV MIEP or the HIV-1 promoter was monitored (as TFIIH consists of nine subunits, and one of them, Cdk7, possesses kinase activity. TFIIH contains a DNA-dependent ATPase activity, two ATP-dependent DNA helicase activities, and a kinase activity specific for the CTD of RNAPII ( Drapkin and Reinberg 1994; TFIIH first came to TFIIH transcriptional activity and TFIIH-associated CTD kinase activity are repressed in the mitotic extract. chemical genetics CTD kinase TFIIH disassembly T FIIH, a 10-subunit complex with many resident enzymatic activities,isessentialfortranscriptionbyRNApolymeraseII (Pol II) (15). Here the substrate specificities of TFIIH and two forms of the Cdk7-containing kinase complex are Concomitant with the open complex Recently, a cyclin-dependent kinase-activator kinase (MO15 and cyclin H) was Cdk7 is the kinase catalytic subunit of CAK, the Cdc2 activating kinase complex, consisting of Cdk7, cyclin H and MAT-1 (19-21). 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