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Eukaryotic Cell, February 2002, p. 95-104, Vol. 1, No. 1
1535-9778/02/$04.00+0     DOI: 10.1128/EC.1.1.95-104.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

CDC42 Is Required for Polarized Growth in Human Pathogen Candida albicans{dagger}

*** Sophia C. Ushinsky,1 Doreen Harcus,1 Josee Ash,1 Daniel Dignard,1 Anne Marcil,1 Joachim Morchhauser,2 David Y. Thomas,1,3,4,{ddagger} Malcolm Whiteway,1,3* and Ekkehard Leberer1,5,§

Eukaryotic Genetics, National Research Council Biotechnology Research Institute, Montreal, Quebec, Canada, H4P 2R2,1 Zentrum für Infektionsforschung, Universität Würzburg, D-97070 Würzburg, Germany,2 Departments of Biology,3 Anatomy and Cell Biology,4 Experimental Medicine, McGill University, Montreal, Quebec, Canada5

Received 1 August 2001/ Accepted 9 November 2001

Cdc42p is a member of the RAS superfamily of GTPases and plays an essential role in polarized growth in many eukaryotic cells. We cloned the Candida albicans CaCDC42 by functional complementation in Saccharomyces cerevisiae and analyzed its function in C. albicans. A double deletion of CaCDC42 was made in a C. albicans strain containing CaCDC42 under the control of the PCK1 promoter. When expression of the heterologous copy of CaCDC42 was repressed in this strain, the cells ceased proliferation. These arrested cells were large, round, and unbudded and contained predominantly two nuclei. The PCK1-mediated overexpression of wild-type CaCdc42p had no effect on cells. However, in cells overexpressing CaCdc42p containing the dominant-negative D118A substitution, proliferation was blocked and the arrested cells were large, round, unbudded, and multinucleated, similar to the phenotype of the cdc42 double-deletion strain. Cells overexpressing CaCdc42p containing the hyperactive G12V substitution also ceased proliferation in yeast growth medium; in this case the arrested cells were multinucleated and multibudded. An intact CAAX box is essential for the phenotypes associated with either CaCdc42pG12V or CaCdc42pD118A ectopic expression, suggesting that membrane attachment is involved in CaCdc42p function. In addition, the lethality caused by ectopic expression of CaCdc42pG12V was suppressed by deletion of CST20 but not by deletion of CaCLA4. CaCdc42p function was also examined under hypha-inducing conditions. Cdc42p depletion prior to hyphal induction trapped cells in a round, unbudded state, while depletion triggered at the same time as hyphal induction permitted the initiation of germ tubes that failed to be extended. Ectopic expression of either the G12V or D118A substitution protein modified hyphal formation in a CAAX box-dependent manner. Thus, CaCdc42p function appears important for polarized growth of both the yeast and hyphal forms of C. albicans.

* Corresponding author. Mailing address: Eukaryotic Genetics, National Research Council Biotechnology Research Institute, 6100 Royalmount Ave., Montreal, Quebec, Canada, H4P 2R2. Phone: (514) 496-6146. Fax: (514) 496-6213. E-mail: malcolm.whiteway{at}nrc.ca.

{dagger} Dedicated to the memory of Teresa Keng. National Research Council of Canada publication 44793.

{ddagger} Present address: Chair of Biochemistry, McGill University, Montreal, Quebec, Canada.

§ Present address: Aventis Genomics Center, Aventis Pharma, D-82152 Martinsried, Germany.

Eukaryotic Cell, February 2002, p. 95-104, Vol. 1, No. 1
1535-9778/02/$04.00+0     DOI: 10.1128/EC.1.1.95-104.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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