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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.
*
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.
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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