Volume 272, Number 20,
Issue of May 16, 1997
pp. 13180-13188
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
Overexpression of the G1-cyclin Gene CLN2
Represses the Mating Pathway in Saccharomyces
cerevisiae at the Level of the MEKK Ste11
(Received for publication, November 15, 1996, and in revised form, March 7, 1997)
Katja
Wassmann
and
Gustav
Ammerer
From the Institute for Biochemistry and Molecular Cell Biology,
Ludwig Boltzmann Forschungsstelle, University of Vienna, Dr. Bohr,
Gasse 9, 1030 Vienna, Austria
Basal and induced transcription of
pheromone-dependent genes is regulated in a cell
cycle-dependent way. FUS1, a gene strongly induced after
pheromone treatment, shows high mRNA levels in mitosis and early
G1 phase of the cell cycle, a decrease in G1
after START and again an increase in S phase. Overexpression of
CLN2 was shown to repress the transcript number of
pheromone-dependent genes (1). We asked whether the
activities of components of the mating pathway fluctuate during the
cell cycle. We were also interested in determining at what level Cln2
represses the signal transduction machinery. Here we show that the
activity of the mitogen-activated protein kinase Fus3 indeed fluctuates
during the cell cycle, reflecting the oscillations of the gene
transcripts. CLN2 overexpression represses Fus3 kinase
activity, independently of the phosphatase Msg5. Additionally, we show
that the activity of the MEK Ste7 also fluctuates during the cell
cycle. Increased Cln2 levels repress the ability of hyperactive
STE11 alleles to induce the pathway. G protein-independent
activation of Ste11 caused by an rga1 pbs2 mutation is
resistant to high levels of Cln2 kinase. Therefore our results suggest
that Cln2-dependent repression of the mating pathway occurs
at the level of Ste11.