TFIIH-Associated Cdk7 Kinase Functions in Phosphorylation of C-Terminal Domain Ser7 Residues, Promoter-Proximal Pausing, and Termination by RNA Polymerase II Export

@article{citeulike:6083582, abstract = {The function of human TFIIH-associated Cdk7 in RNA polymerase II (Pol II) transcription and C-terminal domain (CTD) phosphorylation was investigated in analogue-sensitive Cdk7as/as mutant cells where the kinase can be inhibited without disrupting TFIIH. We show that both Cdk7 and Cdk9/PTEFb contribute to phosphorylation of Pol II CTD Ser5 residues on transcribed genes. Cdk7 is also a major kinase of CTD Ser7 on Pol II at the c-fos and U snRNA genes. Furthermore, TFIIH and recombinant Cdk7-CycH-Mat1 as well as recombinant Cdk9-CycT1 phosphorylated CTD Ser7 and Ser5 residues in vitro. Inhibition of Cdk7 in vivo suppressed the amount of Pol II accumulated at 5' ends on several genes including c-myc, p21, and glyceraldehyde-3-phosphate dehydrogenase genes, indicating reduced promoter-proximal pausing or polymerase "leaking" into the gene. Consistent with a 5' pausing defect, Cdk7 inhibition reduced recruitment of the negative elongation factor NELF at start sites. A role of Cdk7 in regulating elongation is further suggested by enhanced histone H4 acetylation and diminished histone H4 trimethylation on lysine 36--two marks of elongation--within genes when the kinase was inhibited. Consistent with a new role for TFIIH at 3' ends, it was detected within genes and 3'-flanking regions, and Cdk7 inhibition delayed pausing and transcription termination. 10.1128/MCB.00637-09}, author = {Glover-Cutter, Kira and Larochelle, Stephane and Erickson, Benjamin and Zhang, Chao and Shokat, Kevan and Fisher, Robert P. and Bentley, David L.}, citeulike-article-id = {6083582}, citeulike-linkout-0 = {http://dx.doi.org/10.1128/MCB.00637-09}, citeulike-linkout-1 = {http://mcb.asm.org/content/29/20/5455.abstract}, citeulike-linkout-2 = {http://mcb.asm.org/content/29/20/5455.full.pdf}, citeulike-linkout-3 = {http://view.ncbi.nlm.nih.gov/pubmed/19667075}, citeulike-linkout-4 = {http://www.hubmed.org/display.cgi?uids=19667075}, day = {15}, doi = {10.1128/MCB.00637-09}, journal = {Mol. Cell. Biol.}, keywords = {cdk7, d-polymerase}, month = {October}, number = {20}, pages = {5455--5464}, posted-at = {2009-11-08 01:14:17}, priority = {2}, title = {TFIIH-Associated Cdk7 Kinase Functions in Phosphorylation of C-Terminal Domain Ser7 Residues, Promoter-Proximal Pausing, and Termination by RNA Polymerase II}, url = {http://dx.doi.org/10.1128/MCB.00637-09}, volume = {29}, year = {2009} }

TY - JOUR ID - citeulike:6083582 L3 - citeulike-article-id:6083582 N2 - The function of human TFIIH-associated Cdk7 in RNA polymerase II (Pol II) transcription and C-terminal domain (CTD) phosphorylation was investigated in analogue-sensitive Cdk7as/as mutant cells where the kinase can be inhibited without disrupting TFIIH. We show that both Cdk7 and Cdk9/PTEFb contribute to phosphorylation of Pol II CTD Ser5 residues on transcribed genes. Cdk7 is also a major kinase of CTD Ser7 on Pol II at the c-fos and U snRNA genes. Furthermore, TFIIH and recombinant Cdk7-CycH-Mat1 as well as recombinant Cdk9-CycT1 phosphorylated CTD Ser7 and Ser5 residues in vitro. Inhibition of Cdk7 in vivo suppressed the amount of Pol II accumulated at 5' ends on several genes including c-myc, p21, and glyceraldehyde-3-phosphate dehydrogenase genes, indicating reduced promoter-proximal pausing or polymerase "leaking" into the gene. Consistent with a 5' pausing defect, Cdk7 inhibition reduced recruitment of the negative elongation factor NELF at start sites. A role of Cdk7 in regulating elongation is further suggested by enhanced histone H4 acetylation and diminished histone H4 trimethylation on lysine 36--two marks of elongation--within genes when the kinase was inhibited. Consistent with a new role for TFIIH at 3' ends, it was detected within genes and 3'-flanking regions, and Cdk7 inhibition delayed pausing and transcription termination. 10.1128/MCB.00637-09 IS - 20 JF - Mol. Cell. Biol. EP - 5464 TI - TFIIH-Associated Cdk7 Kinase Functions in Phosphorylation of C-Terminal Domain Ser7 Residues, Promoter-Proximal Pausing, and Termination by RNA Polymerase II VL - 29 SP - 5455 KW - cdk7 KW - d-polymerase AU - Glover-Cutter, Kira AU - Larochelle, Stephane AU - Erickson, Benjamin AU - Zhang, Chao AU - Shokat, Kevan AU - Fisher, Robert AU - Bentley, David PY - 2009/10/15/ UR - http://dx.doi.org/10.1128/MCB.00637-09 ER -