Carlsberg Laboratory, Yeast Biology, Valby, Copenhagen, Denmark.
Curr Genet. 2010 Aug;56(4):309-19. doi: 10.1007/s00294-010-0301-7. Epub 2010 Apr 11.
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 Deltasla1 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 Deltasla1 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 Deltanbp2 strains lacked a large vacuole but initiated several germ tubes. The mutant phenotypes of Deltanbp2 and Deltasla1 could be corrected by reintegration of the wild-type genes.
本报告描述了三个编码 Src Homology 3(SH3)结构域蛋白的白色念珠菌基因的分析。酿酒酵母中的同源物由 SLA1、NBP2 和 CYK3 基因编码。在白色念珠菌中删除 CYK3 是不可行的,这表明它是必需的。CaCYK3 的启动子关闭实验显示出胞质分裂缺陷,这与 GFP 标记的 Cyk3 在隔膜部位的定位一致。SLA1 的缺失导致菌株形成菌丝体丝状的能力下降。在所有生长阶段,Deltasla1 菌株中的皮质肌动蛋白斑的数量都大大减少。Sla1-GFP 定位于在菌丝尖端集中发现的斑中。Sla1 的前两个 SH3 结构域的缺失仍导致截短蛋白的皮质定位。然而,该菌株中的肌动蛋白细胞骨架异常,类似于 Deltasla1 缺失突变体,表明这些 SH3 结构域的功能是将肌动蛋白成核募集到内吞作用部位。NBP2 的缺失导致菌丝中液泡融合缺陷。Deltanbp2 菌株的生殖细胞缺乏大液泡,但启动了几个生殖管。Deltanbp2 和 Deltasla1 的突变表型可以通过野生型基因的重新整合来纠正。