Cdk-activating kinase complex is a component of human transcription factor TFIIH

R Shiekhattar; F Mermelstein; R P Fisher; R Drapkin; B Dynlacht; H C Wessling; D O Morgan; D Reinberg

doi:10.1038/374283a0

Cdk-activating kinase complex is a component of human transcription factor TFIIH

Nature. 1995 Mar 16;374(6519):283-7. doi: 10.1038/374283a0.

Authors

R Shiekhattar¹, F Mermelstein, R P Fisher, R Drapkin, B Dynlacht, H C Wessling, D O Morgan, D Reinberg

Affiliation

¹ Howard Hughes Medical Institute, Department of Biochemistry, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway 08854-5635.

PMID: 7533895
DOI: 10.1038/374283a0

Abstract

Transcription factor IIH (TFIIH) contains a kinase capable of phosphorylating the carboxy-terminal domain (CTD) of the largest subunit of RNA polymerase II (RNAPII). Here we report the identification of the Cdk-activating kinase (Cak) complex (Cdk7 and cyclin H) as a component of TFIIH after extensive purification of TFIIH by chromatography. We find that affinity-purified antibodies directed against cyclin H inhibit TFIIH-dependent transcription and that both cyclin H and Cdk7 antibodies inhibit phosphorylation of the CTD of the largest subunit of the RNAPII in the preinitiation complex. Cak is present in at least two distinct complexes, TFIIH and a smaller complex that is unable to phosphorylate RNAPII in the preinitiation complex. Both Cak complexes, as well as recombinant Cak, phosphorylate a CTD peptide. Finally, TFIIH was shown to phosphorylate both Cdc2 and Cdk2, suggesting that there could be a link between transcription and the cell cycle machinery.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

MeSH terms

Amino Acid Sequence
Chromatography, Affinity
Cyclin H
Cyclin-Dependent Kinase-Activating Kinase
Cyclin-Dependent Kinases*
Cyclins / metabolism
Detergents
HeLa Cells
Humans
Immunologic Techniques
Molecular Sequence Data
Phosphorylation
Protein Kinases / metabolism
Protein Serine-Threonine Kinases / isolation & purification
Protein Serine-Threonine Kinases / metabolism*
RNA Polymerase II / metabolism
Recombinant Proteins / metabolism
Sarcosine / analogs & derivatives
Transcription Factor TFIIH
Transcription Factors / chemistry
Transcription Factors / metabolism*
Transcription Factors, TFII*
Transcription, Genetic

Substances

CCNH protein, human
Cyclin H
Cyclins
Detergents
Recombinant Proteins
Transcription Factors
Transcription Factors, TFII
Transcription Factor TFIIH
sarkosyl
Protein Kinases
carboxy-terminal domain kinase
Protein Serine-Threonine Kinases
Cyclin-Dependent Kinases
RNA Polymerase II
Sarcosine
Cyclin-Dependent Kinase-Activating Kinase
CDK7 protein, human