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J. Biol. Chem., Vol. 279, Issue 36, 37512-37517, September 3, 2004

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NPR1 Kinase and RSP5-BUL1/2 Ubiquitin Ligase Control GLN3-dependent Transcription in Saccharomyces cerevisiae^*

José L. Crespo, Stephen B. Helliwell, Christa Wiederkehr¶||, Philippe Demougin¶, Brian Fowler**, Michael Primig¶, and Michael N. Hall

From the Division of Biochemistry and the ^¶Swiss Institute of Bioinformatics, Biozentrum, University of Basel, Klingelbergstrasse 70, CH-4056 Basel, Switzerland and the ^**University Childrens Hospital, CH-4005 Basel, Switzerland

The GATA transcription factors GLN3 and GAT1 activate nitrogen-regulated genes in Saccharomyces cerevisiae. NPR1 is a protein kinase that controls post-Golgi sorting of amino acid permeases. In the presence of a good nitrogen source, TOR (target of rapamycin) maintains GLN3 and NPR1 phosphorylated and inactive by inhibiting the type 2A-related phosphatase SIT4. We identified NPR1 as a regulator of GLN3. Specifically, loss of NPR1 causes nuclear translocation and activation of GLN3, but not GAT1, in nitrogen-rich conditions. NPR1-mediated inhibition of GLN3 is independent of the phosphatase SIT4. We also demonstrate that the E3/E4 ubiquitin-protein ligase proteins RSP5 and BUL1/2 are required for GLN3 activation under poor nitrogen conditions. Thus, NPR1 and BUL1/2 antagonistically control GLN3-dependent transcription, suggesting a role for regulated ubiquitination in the control of nutrient-responsive transcription.

Received for publication, July 1, 2004

^* This work was supported by grants from the Canton of Basel and the Swiss National Science Foundation (to M. N. H.) and the Roche Research Foundation (to S. B. H.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Recipient of a Federation of European Biochemical Societies longterm fellowship.

^|| Supported by the Swiss Institute of Bioinformatics.

To whom correspondence should be addressed. Tel.: 41-61-267-2150, Fax: 41-61-267-2149; E-mail: m.hall{at}unibas.ch.

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