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Mol Cell Biol March 1, 1998; 18 (3): 1467-76.

Repression of TFIIH transcriptional activity and TFIIH-associated cdk7 kinase activity at mitosis.

Long JJ , Leresche A , Kriwacki RW , Gottesfeld JM .

Nuclear transcription is repressed when eukaryotic cells enter mitosis. Mitotic repression of transcription of various cellular and viral gene promoters by RNA polymerase II can be reproduced in vitro either with extracts prepared from cells arrested at mitosis with the microtubule polymerization inhibitor nocodazole or with nuclear extracts prepared from asynchronous cells and the mitotic protein kinase cdc2/cyclin B. Purified cdc2/cyclin B kinase is also sufficient to inhibit transcription in reconstituted transcription reactions with biochemically purified and recombinant basal transcription factors and RNA polymerase II. The cyclin-dependent kinase inhibitor p21Waf1/Cip1/Sdi1 can reverse the effect of cdc2/cyclin B kinase, indicating that repression of transcription is due to protein phosphorylation. Transcription rescue and inhibition experiments with each of the basal factors and the polymerase suggest that multiple components of the transcription machinery are inactivated by cdc2/cyclin B kinase. 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. Protein labeling experiments indicate that the p62 and p36 subunits of TFIIH are in vitro substrates for mitotic phosphorylation. Using the carboxy-terminal domain of the large subunit of RNA polymerase II as a test substrate for phosphorylation, the TFIIH-associated kinase, cdk7/cyclin H, is inhibited concomitant with inhibition of transcription activity. Our results suggest that there exist multiple phosphorylation targets for the global shutdown of transcription at mitosis.

PubMed ID: 9488463
PMC ID: PMC108861
Article link: Mol Cell Biol
Grant support: [+]

Species referenced: Xenopus
Genes referenced: anxa2 cdk7 gtf2h4 nup62 sqstm1 taf6 tbp

References [+] :
Adamczewski, MAT1, cdk7 and cyclin H form a kinase complex which is UV light-sensitive upon association with TFIIH. 1996, Pubmed