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Eukaryotic Cell, October 2002, p. 663-672, Vol. 1, No. 5
1535-9778/02/$04.00+0     DOI: 10.1128/EC.1.5.663-672.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Amino Acid-Dependent Gcn4p Stability Regulation Occurs Exclusively in the Yeast Nucleus

Ralph Pries, Katrin Bömeke, Stefan Irniger, Olav Grundmann, and Gerhard H. Braus*

Institute of Microbiology and Genetics, Georg-August-University, D-37077 Göttingen, Germany

Received 26 December 2001/ Accepted 12 June 2002

The c-Jun-like transcriptional activator Gcn4p controls biosynthesis of translational precursors in the yeast Saccharomyces cerevisiae. Protein stability is dependent on amino acid limitation and cis signals within Gcn4p which are recognized by cyclin-dependent protein kinases, including Pho85p. The Gcn4p population within unstarved yeast consists of a small relatively stable cytoplasmic fraction and a larger less stable nuclear fraction. Gcn4p contains two nuclear localization signals (NLS) which function independently of the presence or absence of amino acids. Expression of NLS-truncated Gcn4p results in an increased cytoplasmic fraction and an overall stabilization of the protein. The same effect is achieved for the entire Gcn4p in a yrb1 yeast mutant strain impaired in the nuclear import machinery. In the presence of amino acids, controlled destabilization of Gcn4p is triggered by the phosphorylation activity of Pho85p. A pho85{Delta} mutation stabilizes Gcn4p without affecting nuclear import. Pho85p is localized within the nucleus in the presence or absence of amino acids. Therefore, there is a strict spatial separation of protein synthesis and degradation of Gcn4p in yeast. Control of protein stabilization which antagonizes Gcn4p function is restricted to the nucleus.


* Corresponding author. Mailing address: Institute of Microbiology and Genetics, Georg-August-University, Grisebachstrasse 8, D-37077 Göttingen, Germany. Phone: 49-551-393771. Fax: 49-551-393820. E-mail: gbraus{at}gwdg.de.


Eukaryotic Cell, October 2002, p. 663-672, Vol. 1, No. 5
1535-9778/02/$04.00+0     DOI: 10.1128/EC.1.5.663-672.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.




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