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Molecular and Cellular Biology, December 1999, p. 8042-8051, Vol. 19, No. 12
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

A Subunit of Yeast TFIIIC Participates in the Recruitment of TATA-Binding Protein

Eric Deprez, Rosalía Arrebola, Christine Conesa, and André Sentenac^*

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

Received 29 June 1999/Returned for modification 5 August 1999/Accepted 7 September 1999

TFIIIC plays a key role in nucleating the assembly of the initiation factor TFIIIB on class III genes. We have characterized an essential gene, TFC8, encoding the 60-kDa polypeptide, 60, present in affinity-purified TFIIIC. Hemagglutinin-tagged variants of 60 were found to be part of TFIIIC-tDNA complexes and to reside at least in part in the downstream DNA-binding domain B. Unexpectedly, the thermosensitive phenotype of N-terminally tagged 60 was suppressed by overexpression of 95, which belongs to the A domain, and by two TFIIIB components, TATA-binding protein (TBP) and B"/TFIIIB90 (but not by TFIIIB70). Mutant TFIIIC was deficient in the activation of certain tRNA genes in vitro, and the transcription defect was selectively alleviated by increasing TBP concentration. Coimmunoprecipitation experiments support a direct interaction between TBP and 60. It is suggested that 60 links A and B domains and participates in TFIIIB assembly via its interaction with TBP.

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^* Corresponding author. Mailing address: Service de Biochimie et de Génétique Moléculaire, CEA/Saclay, F-91191 Gif-sur-Yvette, France. Phone: 33-1-69-08-22-36. Fax: 33-1-69-08-47-12. E-mail: sentenac{at}dsvidf.cea.fr.

Present address: Laboratoire de Physicochimie et Pharmacologie des Macromolécules Biologiques (UMR8532), ENS de Cachan, F-94235 Cachan Cedex, France.

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Molecular and Cellular Biology, December 1999, p. 8042-8051, Vol. 19, No. 12
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

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