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Papers In Press, published online ahead of print January 16, 2003
J. Biol. Chem, 10.1074/jbc.M213310200
Submitted on December 9, 2002
Revised on January 16, 2003
Accepted on January 16, 2003

The tau 95 subunit of yeast TFIIIC influences upstream and downstream functions of TFIIIC5dna complexes

Sabine Jourdain, Joël Acker, Cécile Ducrot, André Sentenac, and Olivier ;Lefebvre

DBJC, CEA, Gif-Sur-Yvette F-91191

Corresponding Author: Olivier.Lefebvre{at}Cea.Fr

The yeast transcription factor IIIC (TFIIIC) is organized in two distinct, multisubunit domains, tau A and tau B, which 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 B5DNA 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 TFIIIC5DNA complexes and start site selection. Mutant TFIIIC5DNA complexes presented a shift in their 5' border, generated slow-migrating TFIIIB5DNA complexes upon stripping TFIIIC by heparin or heat treatment, and allowed initiation at downstream sites. In addition, mutant TFIIIC5DNA 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 A5DNA spatial configuration and tau B5DNA stability.


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