Mechanism of Promoter Melting by the Xeroderma Pigmentosum Complementation Group B Helicase of Transcription Factor IIH Revealed by Protein-DNA Photo-Cross-Linking

doi:10.1128/MCB.20.21.8168-8177.2000

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Molecular and Cellular Biology, November 2000, p. 8168-8177, Vol. 20, No. 21
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Mechanism of Promoter Melting by the Xeroderma Pigmentosum Complementation Group B Helicase of Transcription Factor IIH Revealed by Protein-DNA Photo-Cross-Linking

Maxime Douziech,¹ Frédéric Coin,² Jean-Marc Chipoulet,² Yoko Arai,³ Yoshiaki Ohkuma,³ Jean-Marc Egly,² and Benoit Coulombe¹^,^*

Département de Biologie, Faculté des Sciences, Université de Sherbrooke, Sherbrooke, Québec J1K 2R1, Canada¹; Institut de Génétique et de Biologie Moléculaire et Cellulaire, UPR 6520 (CNRS), Unité 184 (INSERM), Illkirch Cédex, CU de Strasbourg, France²; and Institute for Molecular and Cellular Biology, Institute for Molecular and Cellular Biology and Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka 565-0871, Japan³

Received 23 June 2000/Returned for modification 18 July 2000/Accepted 26 July 2000

The p89/xeroderma pigmentosum complementation group B (XPB) ATPase-helicase of transcription factor IIH (TFIIH) is essentialfor promoter melting prior to transcription initiation by RNApolymerase II (RNAPII). By studying the topological organizationof the initiation complex using site-specific protein-DNA photo-cross-linking,we have shown that p89/XPB makes promoter contacts both upstreamand downstream of the initiation site. The upstream contact, whichis in the region where promoter melting occurs (positions $-$ 9 to+2), requires tight DNA wrapping around RNAPII. The addition ofhydrolyzable ATP tethers the template strand at positions $-$ 5 and+1 to RNAPII subunits. A mutation in p89/XPB found in a xerodermapigmentosum patient impairs the ability of TFIIH to associatecorrectly with the complex and thereby melt promoter DNA. A modelfor open complex formation isproposed.

^* Corresponding author. Mailing address: Département de Biologie, Faculté des Sciences, Université de Sherbrooke, Sherbrooke,Québec J1K 2R1, Canada. Phone: (819) 821-8000, ext. 1092. Fax:(819) 821-7083. E-mail: b.coulom{at}courrier.usherb.ca.

Molecular and Cellular Biology, November 2000, p. 8168-8177, Vol. 20, No. 21
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

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