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Microbiology 148 (2002), 4035-4048
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Microbiology (2002), 148, 4035-4048.
© 2002 Society for General Microbiology

Research Paper

The protein kinase Kic1 affects 1,6-ß-glucan levels in the cell wall of Saccharomyces cerevisiae

Edwin Vinke,a,1, Jack H. Vossene,b,1, Arthur F. J. Ramc,1, Herman van den Ende1, Stephan Brekelmans1, Hans de Nobeld,1 and Frans M. Klis1

Swammerdam Institute for Life Sciences, University of Amsterdam, Nieuwe Achtergracht 166, 1018 WV, Amsterdam, The Netherlands1

Author for correspondence: Frans M. Klis. Tel: +31 20 525 7834. Fax: +31 20 525 7056. e-mail: klis{at}science.uva.nl

KIC1 encodes a PAK kinase that is involved in morphogenesis and cell integrity. Both over- and underexpressing conditions of KIC1 affected cell wall composition. Kic1-deficient cells were hypersensitive to the cell wall perturbing agent calcofluor white and had less 1,6-ß-glucan. When Kic1-deficient cells were crossed with various kre mutants, which also have less 1,6-ß-glucan in their wall, the double mutants displayed synthetic growth defects. However, when crossed with the 1,3-ß-glucan-deficient strain fks1{Delta}, no synthetic growth defect was observed, supporting a specific role for KIC1 in regulating 1,6-ß-glucan levels. Kic1-deficient cells also became highly resistant to the cell wall-degrading enzyme mixture Zymolyase, and exhibited higher transcript levels of the cell wall protein-encoding genes CWP2 and SED1. Conversely, overexpression of KIC1 resulted in increased sensitivity to Zymolyase and in a higher level of 1,6-ß-glucan. Multicopy suppressor analysis of a Kic1-deficient strain identified RHO3. Consistent with this, expression levels of RHO3 correlated with 1,6-ß-glucan levels in the cell wall. Interestingly, expression levels of KIC1 and the MAP kinase kinase PBS2 had opposite effects on Zymolyase sensitivity of the cells and on cell wall 1,6-ß-glucan levels in the wall. It is proposed that Kic1 affects cell wall construction in multiple ways and in particular in regulating 1,6-ß-glucan levels in the wall.

Keywords: KIC1/CWH30, PBS2, RHO3, cell wall synthesis

Abbreviations: HOG, high osmolarity glycerol response; MAPK, mitogen-activated protein kinase; SPB, spindle pole body

e These authors contributed equally to this work.

a Present address: AMC Liver Center, Meibergdreef 69-71, 1105 BK, Amsterdam, The Netherlands.

b Present address: Section of Plant Pathology, Swammerdam Institute for Life Sciences, University of Amsterdam, Kruislaan 318, 1098 SM, Amsterdam, The Netherlands.

c Present address: Institute of Molecular Plant Sciences, Leiden University, Wassenaarseweg 64, 2333 AL, Leiden, The Netherlands.

d Present address: Genencor International B. V., PO Box 218, 2300 AE, Leiden, The Netherlands.




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