Zhang Bing, Yu Qilin, Jia Chang, Wang Yuzhou, Xiao Chenpeng, Dong Yijie, Xu Ning, Wang Lei, Li Mingchun
Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Department of Microbiology, Nankai University, Tianjin, PR China.
Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Department of Microbiology, Nankai University, Tianjin, PR China; Nankai University, TEDA, Sch Biol Sci & Biotechnol, Tianjin, PR China.
Fungal Genet Biol. 2015 Aug;81:261-70. doi: 10.1016/j.fgb.2014.12.007. Epub 2015 Jan 6.
Candida albicans is a common pathogenic fungus and has aroused widespread attention recently. Actin cytoskeleton, an important player in polarized growth, protein secretion and organization of cell shape, displays irreplaceable role in hyphal development and cell integrity. In this study, we demonstrated a homologue of Saccharomyces cerevisiae Sac1, in C. albicans. It is a potential PIP phosphatase with Sac domain which is related to actin organization, hyphal development, biofilm formation and cell wall integrity. Deletion of SAC1 did not lead to insitiol-auxotroph phenotype in C. albicans, but this gene rescued the growth defect of S. cerevisiae sac1Δ in the insitiol-free medium. Hyphal induction further revealed the deficiency of sac1Δ/Δ in hyphal development and biofilm formation. Fluorescence observation and real time PCR (RT-PCR) analysis suggested both actin and the hyphal cell wall protein Hwp1 were overexpressed and mislocated in this mutant. Furthermore, cell wall integrity (CWI) was largely affected by deletion of SAC1, due to the hypersensitivity to cell wall stress, changed content and distribution of chitin in the mutant. As a result, the virulence of sac1Δ/Δ was seriously attenuated. Taken together, this study provides evidence that Sac1, as a potential PIP phosphatase, is essential for actin organization, hyphal development, CWI and pathogenicity in C. albicans.
白色念珠菌是一种常见的致病真菌,近年来引起了广泛关注。肌动蛋白细胞骨架在极化生长、蛋白质分泌和细胞形态组织中起着重要作用,在菌丝发育和细胞完整性方面发挥着不可替代的作用。在本研究中,我们在白色念珠菌中证实了酿酒酵母Sac1的一个同源物。它是一种潜在的具有Sac结构域的磷脂酰肌醇磷酸酶,与肌动蛋白组织、菌丝发育、生物膜形成和细胞壁完整性有关。缺失SAC1不会导致白色念珠菌出现肌醇营养缺陷型表型,但该基因挽救了酿酒酵母sac1Δ在无肌醇培养基中的生长缺陷。菌丝诱导进一步揭示了sac1Δ/Δ在菌丝发育和生物膜形成方面的缺陷。荧光观察和实时PCR(RT-PCR)分析表明,肌动蛋白和菌丝细胞壁蛋白Hwp1在该突变体中均过度表达且定位错误。此外,由于对细胞壁应激的超敏反应、突变体中几丁质含量和分布的改变,SAC1的缺失在很大程度上影响了细胞壁完整性(CWI)。结果,sac1Δ/Δ的毒力严重减弱。综上所述,本研究提供了证据表明Sac1作为一种潜在的磷脂酰肌醇磷酸酶,对于白色念珠菌的肌动蛋白组织、菌丝发育、CWI和致病性至关重要。