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Molecular and Cellular Biology, January 2003, p. 229-237, Vol. 23, No. 1
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.1.229-237.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Targeting the MEF2-Like Transcription Factor Smp1 by the Stress-Activated Hog1 Mitogen-Activated Protein Kinase

Eulàlia de Nadal, Laura Casadomé, and Francesc Posas*

Cell Signaling Unit, Departament de Ciències Experimentals i de la Salut, Universitat Pompeu Fabra, Barcelona E-08003, Spain

Received 21 June 2002/ Returned for modification 5 August 2002/ Accepted 2 October 2002

Exposure of Saccharomyces cerevisiae to increases in extracellular osmolarity activates the stress-activated Hog1 mitogen-activated protein kinase (MAPK), which is essential for cell survival upon osmotic stress. Yeast cells respond to osmotic stress by inducing the expression of a very large number of genes, and the Hog1 MAPK plays a critical role in gene transcription upon stress. To understand how Hog1 controls gene expression, we designed a genetic screen to isolate new transcription factors under the control of the MAPK and identified the MEF2-like transcription factor, Smp1, as a target for Hog1. Overexpression of SMP1 induced Hog1-dependent expression of osmoresponsive genes such as STL1, whereas smp1{Delta} cells were defective in their expression. Consistently, smp1{Delta} cells displayed reduced viability upon osmotic shock. In vivo coprecipitation and phosphorylation studies showed that Smp1 and Hog1 interact and that Smp1 is phosphorylated upon osmotic stress in a Hog1-dependent manner. Hog1 phosphorylated Smp1 in vitro at the C-terminal region. Phosphorylation of Smp1 by the MAPK is essential for its function, since a mutant allele unable to be phosphorylated by the MAPK displays impaired stress responses. Thus, our data indicate that Smp1 acts downstream of Hog1, controlling a subset of the responses induced by the MAPK. Moreover, Smp1 concentrates in the nucleus during the stationary phase, and the lack of SMP1 results in cells that lose viability in the stationary phase. Localization of Smp1 depends on HOG1, and consistently, hog1{Delta} cells also lose viability during this growth phase. These data suggest that Smp1 could be mediating a role for the Hog1 MAPK during the stationary phase.

* Corresponding author. Mailing address: Cell Signaling Unit, Facultat de Ciències de la Salut i de la Vida, Universitat Pompeu Fabra (UPF), C/Doctor Aiguader 80, Barcelona E-08003, Spain. Phone: 34-93-542 2848. Fax: 34-93-542 2802. E-mail: francesc.posas{at}cexs.upf.es.

Molecular and Cellular Biology, January 2003, p. 229-237, Vol. 23, No. 1
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.1.229-237.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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