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MBC in Press, published online ahead of print November 8, 2006
Mol. Biol. Cell 10.1091/mbc.E06-05-0411

A more recent version of this article appeared on January 1, 2007
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Submitted on May 11, 2006
Revised on October 20, 2006
Accepted on October 26, 2006

Regulation of Cdc42 GTPase Activity in the Formation of Hyphae in Candida albicans

Helen Court and Peter Sudbery

Department of Molecular Biology and Biotechnology, Sheffield University, Sheffield S10 2TN, United Kingdom

Monitoring Editor: Daniel Lew

The human fungal pathogen, Candida albicans, can switch between yeast, pseudohyphal and hyphal morphologies. To investigate whether the distinctive characteristics of hyphae are due to increased activity of the Cdc42 GTPase, strains lacking negative regulators of Cdc42 were constructed. Unexpectedly, the deletion of the Cdc42 Rho guanine-dissociation inhibitor, RDI1, resulted in reduced rather than enhanced polarized growth. However, when cells lacking both Cdc42 GTPase activating proteins (GAPs), encoded by RGA2 and BEM3, were grown under pseudohyphal-promoting conditions the bud was highly elongated and lacked a constriction at its base, so that its shape resembled a hyphal germ tube. Moreover, a Spitzenkörper was present at the bud tip, a band of disorganised septin was present at bud base, true septin rings formed within the bud, and nuclei migrated out of the mother cell before the first mitosis. These are all characteristic features of a hyphal germ tube. Intriguingly, we observed hyphal-specific phosphorylation of Rga2 suggesting a possible mechanism for Cdc42 activation during normal hyphal development. In contrast, expression of Cdc42G12V, which is constitutively GTP-bound since it lacks GTPase activity, resulted in swollen cells with prominent and stable septin bars. These results suggest the development of hyphal-specific characteristics is promoted by Cdc42-GTP in a process that also requires the intrinsic GTPase activity of Cdc42.

Address correspondence to: Peter Sudbery (P.Sudbery{at}shef.ac.uk)




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