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Molecular and Cellular Biology, January 2002, p. 298-308, Vol. 22, No. 1
0270-7306/01/$04.00+0     DOI: 10.1128/MCB.22.1.298-308.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Multiple Roles of the {tau}131 Subunit of Yeast Transcription Factor IIIC (TFIIIC) in TFIIIB Assembly

Hélène Dumay-Odelot,,{dagger} Joël Acker, Rosalia Arrebola, André Sentenac, and Christian Marck*

Service de Biochimie et de Génétique Moléculaire, CEA/Saclay, F-91191 Gif-sur-Yvette Cedex, France

Received 22 June 2001/ Returned for modification 3 August 2001/ Accepted 5 October 2001

Yeast transcription factor IIIC (TFIIIC) plays a key role in assembling the transcription initiation factor TFIIIB on class III genes after TFIIIC-DNA binding. The second largest subunit of TFIIIC, {tau}131, is thought to initiate TFIIIB assembly by interacting with Brf1/TFIIIB70. In this work, we have analyzed a TFIIIC mutant ({tau}131-{Delta}TPR2) harboring a deletion in {tau}131 removing the second of its 11 tetratricopeptide repeats. Remarkably, this thermosensitive mutation was selectively suppressed in vivo by overexpression of B"/TFIIIB90, but not Brf1 or TATA-binding protein. In vitro, the mutant factor preincubated at restrictive temperature bound DNA efficiently but lost transcription factor activity. The in vitro transcription defect was abolished at high concentrations of B" but not Brf1. Copurification experiments of baculovirus-expressed proteins confirmed a direct physical interaction between {tau}131 and B". {tau}131, therefore, appears to be involved in the recruitment of both Brf1 and B".

* Corresponding author. Mailing address: Service de Biochimie et de Génétique Moléculaire, CEA/Saclay, F-91191 Gif-sur-Yvette Cedex, France. Phone: 33 1 69 08 46 20. Fax: 33 1 69 08 47 12. E-mail: marck{at}jonas.saclay.cea.fr.

{dagger} Present address: CNRS, UMR 8532, LBPA, École Normale Supérieure de Cachan, F-94235 Cachan Cedex, France.

Molecular and Cellular Biology, January 2002, p. 298-308, Vol. 22, No. 1
0022-538X/01/$04.00+0     DOI: 10.1128/MCB.22.1.298-308.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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