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Originally published In Press as doi:10.1074/jbc.M213310200 on January 16, 2003

J. Biol. Chem., Vol. 278, Issue 12, 10450-10457, March 21, 2003
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The tau 95 Subunit of Yeast TFIIIC Influences Upstream and Downstream Functions of TFIIIC·DNA Complexes*

Sabine JourdainDagger , Joël Acker, Cécile Ducrot, André Sentenac, and Olivier Lefebvre§

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

The yeast transcription factor IIIC (TFIIIC) is organized in two distinct multisubunit domains, tau A and tau B, that are respectively responsible for TFIIIB assembly and stable anchoring of TFIIIC on the B block of tRNA genes. Surprisingly, we found that the removal of tau A by mild proteolysis stabilizes the residual tau B·DNA complexes at high temperatures. Focusing on the well conserved tau 95 subunit that belongs to the tau A domain, we found that the tau 95-E447K mutation has long distance effects on the stability of TFIIIC·DNA complexes and start site selection. Mutant TFIIIC·DNA complexes presented a shift in their 5' border, generated slow-migrating TFIIIB·DNA complexes upon stripping TFIIIC by heparin or heat treatment, and allowed initiation at downstream sites. In addition, mutant TFIIIC·DNA complexes were highly unstable at high temperatures. Coimmunoprecipitation experiments indicated that tau 95 participates in the interconnection of tau A with tau B via its contacts with tau 138 and tau 91 polypeptides. The results suggest that tau 95 serves as a scaffold critical for tau A·DNA spatial configuration and tau B·DNA stability.

* This work was supported by the Fondation pour la Recherche Médicale Fellowship FDT20000915040 (to S. J.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger Present address: Neurodegeneration Research, Neurology-CEDD, GlaxoSmithKline, Third Ave., Harlow, Essex CM19 5AW, UK.

§ To whom correspondence should be addressed: Service de Biochimie et Génétique Moléculaire, Bâtiment 144, CEA/Saclay, F-91191 Gif-sur-Yvette Cedex, France. Tel.: 33-1-69-08-59-57; Fax: 33-1-69-08-47-12; E-mail: Olivier.Lefebvre@Cea.Fr.

Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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