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Molecular and Cellular Biology, May 2001, p. 3096-3104, Vol. 21, No. 9
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.9.3096-3104.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

High-Mobility-Group Proteins NHP6A and NHP6B Participate in Activation of the RNA Polymerase III SNR6 Gene

Sébastien Lopez,1 Magda Livingstone-Zatchej,2 Sabine Jourdain,1 Fritz Thoma,2 André Sentenac,1 and Marie-Claude Marsolier1,*

Service de Biochimie et de Génétique Moléculaire, CEA/Saclay, 91191 Gif-sur-Yvette Cedex, France,1 and Institut für Zellbiologie, Eidgenössische Technische Hochschule, ETH-Hönggerberg, CH-8093 Zurich, Switzerland2

Received 20 December 2000/Accepted 13 February 2001

Transcription of yeast class III genes involves the formation of a transcription initiation complex that comprises RNA polymerase III (Pol III) and the general transcription factors TFIIIB and TFIIIC. Using a genetic screen for positive regulators able to compensate for a deficiency in a promoter element of the SNR6 gene, we isolated the NHP6A and NHP6B genes. Here we show that the high-mobility-group proteins NHP6A and NHP6B are required for the efficient transcription of the SNR6 gene both in vivo and in vitro. The transcripts of wild-type and promoter-defective SNR6 genes decreased or became undetectable in an nhp6ADelta nhp6BDelta double-mutant strain, and the protection over the TATA box of the wild-type SNR6 gene was lost in nhp6ADelta nhp6BDelta cells at 37°C. In vitro, NHP6B specifically stimulated the transcription of SNR6 templates up to fivefold in transcription assays using either cell nuclear extracts from nhp6ADelta nhp6BDelta cells or reconstituted transcription systems. Finally, NHP6B activated SNR6 transcription in a TFIIIC-independent assay. These results indicate that besides the general transcription factors TFIIIB and TFIIIC, additional auxilliary factors are required for the optimal transcription of at least some specific Pol III genes.

* Corresponding author. Mailing address: Service de Biochimie et de Génétique Moléculaire (Bat. 142), CEA/Saclay, F-91191 Gif-sur-Yvette Cedex, France. Phone: 33-1-69-08-83-54. Fax: 33-1-69-08-47-12. E-mail: marsolie{at}jonas.saclay.cea.fr.

Molecular and Cellular Biology, May 2001, p. 3096-3104, Vol. 21, No. 9
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.9.3096-3104.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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