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The Candida albicans AAA ATPase homologue of Saccharomyces cerevisiae Rix7p (YLL034c) is essential for proper morphology, biofilm formation and activity of secreted aspartyl proteinases
A.S.A. Melo1, A.C.B. Padovan1, R.C. Serafim1, L. Puzer2, A.K. Carmona2,
L. Juliano Neto2, A. Brunstein1 and M.R.S. Briones1
1Departamento de Microbiologia, Imunologia e Parasitologia,
2Departamento de Bioquímica, Universidade Federal de São Paulo, São Paulo, SP, Brasil
Corresponding author: M.R.S. Briones
Genet. Mol. Res. 5 (4): 664-687 (2006)
Received June 26, 2006
Accepted September 4, 2006
Published November 9, 2006

ABSTRACT. Proper morphology is essential for the ability of Candida albicans to switch between yeast and hyphae and thereby sustain its virulence. Here we identified, by differential screening, a novel C. albicans AAA ATPase encoding gene, CaYLL34 (RIX7), with enhanced expression in hyphae. Phylogenetic analysis suggests that CaYLL34 belongs to a “VCP-like” subgroup of AAA ATPases essential for yeast viability and contains a bipartite nuclear localization signal. Inactivation of one copy of CaYLL34, by the URA-Blaster method, generated the heterozygous mutant strain M61. This strain has severe phenotypic alterations, such as a highly increased vacuole, abnormal cell shape and reduced growth in different conditions. Also, major pathogenicity factors are affected in M61, for instance, a significant decrease of hypha formation (>90%), surface biofilm adhesion (86%) and secreted aspartyl proteinase activity (76.5%). Our results show that the partial impairment of CaYll34p cellular levels is sufficient to affect the proper cellular morphology and pathogenicity factors and suggest that this protein is required for biogenesis of ribosomal subunits. Accordingly, we propose that the product of CaYLL34 could be tested as a novel target for antifungal drugs.

Key words: AAA ATPases, Candida albicans, CottonPrep, YLL34, RIX7, Differential screening, Insertional mutagenesis


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