This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplemental material
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Google Scholar
Right arrow Articles by Glover-Cutter, K.
Right arrow Articles by Bentley, D. L.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Glover-Cutter, K.
Right arrow Articles by Bentley, D. L.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, October 2009, p. 5455-5464, Vol. 29, No. 20
0270-7306/09/$08.00+0     doi:10.1128/MCB.00637-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

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

Kira Glover-Cutter,1 Stéphane Larochelle,2 Benjamin Erickson,1 Chao Zhang,3 Kevan Shokat,3 Robert P. Fisher,2 and David L. Bentley1*

Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, UCHSC, MS 8101, P.O. Box 6511, Aurora, Colorado 80045,1 Department of Structural and Chemical Biology, Mount Sinai School of Medicine, New York, New York 10065,2 HHMI and Department of Cellular and Molecular Pharmacology, UCSF, San Francisco, California 941433

Received 17 May 2009/ Returned for modification 2 July 2009/ Accepted 3 August 2009

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.

* Corresponding author. Mailing address: Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, UCHSC, MS 8101, P.O. Box 6511, Aurora, CO 80045. Phone: (303) 724-3237. Fax: (303) 724-3215. E-mail: david.bentley{at}UCHSC.edu

{triangledown} Published ahead of print on 10 August 2009.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

Molecular and Cellular Biology, October 2009, p. 5455-5464, Vol. 29, No. 20
0270-7306/09/$08.00+0     doi:10.1128/MCB.00637-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»