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doi:10.1534/genetics.106.058172
A more recent version of this article appeared on June 1, 2006.
REGULAR RESEARCH PAPERS |
The Saccharomyces cerevisiae 14-3-3 proteins are required for the G1/S transition, actin cytoskeleton organization and cell wall integrity
Francisca Lottersberger 1, Andrea Panza 1, Giovanna Lucchini 1, Simonetta Piatti 1 and Maria Pia Longhese 1*
1 Dipartimento di Biotecnologie e Bioscienze, Università di Milano-Bicocca
* To whom correspondence should be addressed. E-mail: mariapia.longhese{at}unimib.it.
Submitted on March 14, 2006
Revised on March 30, 2006
Accepted on 30 March 2006
14-3-3 proteins are highly conserved polypeptides participating in many biological processes by binding phosphorylated target proteins. In S. cerevisiae, two functionally redundant 14-3-3 isoforms are encoded by the BMH1 and BMH2 genes, whose concomitant deletion is lethal. To gain insights into the essential function(s) shared by these proteins, we searched for high dosage suppressors of the growth defects of temperature-sensitive bmh mutants. Both the protein kinase C1 (Pkc1) and its upstream regulators Wsc2 and Mid2 were found as high dosage suppressors of bmh mutants' temperature-sensitivity, indicating a functional interaction between 14-3-3 and Pkc1. Consistent with a role of 14-3-3 proteins in Pkc1-dependent cellular processes, bmh mutants turned out to be severely impaired at restrictive temperature in initiation of DNA replication, polarization of the actin cytoskeleton and budding, as well as in cell wall integrity. Since Pkc1 acts in concert with the Swi4-Swi6 (SBF) transcriptional activator to control all these processes, the defective G1/S transition of bmh might be linked to impaired SBF activity. Consistently, the levels of the G1 cyclin CLN2 transcripts, which are positively regulated by SBF, were dramatically reduced in bmh mutants. Remarkably, budding and DNA replication defects of bmh mutants were suppressed by CLN2 expression from an SBF-independent promoter, suggesting that 14-3-3 proteins could contribute to regulating the late G1 transcriptional program.
Key Words: 14-3-3, G1/S transition, PKC, actin cytoskeleton, cell wall integrity
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