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Cell Press, Molecular Cell, 1(3), p. 87-95, 1999

DOI: 10.1016/s1097-2765(00)80177-x

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Reconstitution of the Transcription Factor TFIIH

Journal article published in 1999 by Franck Tirode ORCID, Didier Busso, Frédéric Coin, Jean-Marc Egly
This paper is available in a repository.
This paper is available in a repository.

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Abstract

To understand the initiation of the transcription of protein-coding genes, we have dissected the role of the basal transcription/DNA repair factor TFIIH. Having succeeded in reconstituting a functionally active TFIIH from baculovirus recombinant polypeptides, we were able to analyze the role of XPB, XPD, and cdk7 subunits in the transcription reaction. Designing mutated recombinant subunits, we show that the XPB helicase is absolutely required for transcription to open the promoter around the start site whereas the XPD helicase, which is dispensable, stimulates transcription and allows the CAK complex to be anchored to TFIIH. In addition, we also show that cdk7 may phosphorylate the carboxy-terminal domain (CTD) of RNA pol II in the absence of promoter opening.