Abstract
This report describes the analyses of three Candida albicans genes that encode Src Homology 3 (SH3)-domain proteins. Homologs in Saccharomyces cerevisiae are encoded by the SLA1, NBP2, and CYK3 genes. Deletion of CYK3 in C. albicans was not feasible, suggesting it is essential. Promoter shutdown experiments of CaCYK3 revealed cytokinesis defects, which are in line with the localization of GFP-tagged Cyk3 at septal sites. Deletion of SLA1 resulted in strains with decreased ability to form hyphal filaments. The number of cortical actin patches was strongly reduced in Δsla1 strains during all growth stages. Sla1-GFP localizes in patches that are found concentrated at the hyphal tip. Deletion of the first two SH3-domains of Sla1 still resulted in cortical localization of the truncated protein. However, the actin cytoskeleton in this strain was aberrant like in the Δsla1 deletion mutant indicating a function of these SH3 domains to recruit actin nucleation to sites of endocytosis. Deletion of NBP2 resulted in a defect in vacuolar fusion in hyphae. Germ cells of Δnbp2 strains lacked a large vacuole but initiated several germ tubes. The mutant phenotypes of Δnbp2 and Δsla1 could be corrected by reintegration of the wild-type genes.
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Acknowledgments
We thank Alexander Johnson, Suzanne Noble, and Aaron Mitchell for generously providing reagents used in this study; Sidsel Ehlers for providing technical assistance and Andrea Walther for support on microscopy. This study was funded by the EU-Marie Curie Research Training Network “Penelope” and we thank members of this consortium for discussions.
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Communicated by C. D'Enfert.
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Reijnst, P., Jorde, S. & Wendland, J. Candida albicans SH3-domain proteins involved in hyphal growth, cytokinesis, and vacuolar morphology. Curr Genet 56, 309–319 (2010). https://doi.org/10.1007/s00294-010-0301-7
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DOI: https://doi.org/10.1007/s00294-010-0301-7