Carbó Natalia, Pérez-Martín José
Departamento de Biotecnología Microbiana, Centro Nacional de Biotecnología CSIC, Madrid, Spain.
Fungal Genet Biol. 2008 Sep;45(9):1315-27. doi: 10.1016/j.fgb.2008.06.010. Epub 2008 Jul 11.
The increasing evidence linking regulation of polar growth and pathogenicity in fungi has elicited a significant effort devoted to produce a better understanding of mechanisms determining polarization in pathogenic fungi. Here we characterize in the phytopathogenic basidiomycete Ustilago maydis, the Spa2 protein, a well-known component of polarisome, firstly described in Saccharomyces cerevisiae. U. maydis display a dimorphic switch between budding growth of hapoid cells and filamentous growth of the dikaryon. During yeast growth, a GFP-tagged Spa2 protein localized to distinct growth sites in a cell cycle-specific manner, while during hyphal growth is persistently located to hyphal tips. Deletion of spa2 gene produces rounder budding cells and thicker filaments than wild-type cells, suggesting a role of Spa2 for the determination of the growth area in U. maydis. We also address the connections between Spa2 and the actin- and microtubule-cytoskeleton. We found that the absence of Spa2 does not affect cytoskeleton organization and strikingly, interference with actin filament or microtubule formation does not affect the polar localization of Spa2. In contrast, defects in the small GTPase Rac1 seems to affect the ability of Spa2 to locate to precise sites at the tip cell. Finally, to our surprise, we found that cells defectives in Spa2 function were as pathogenic as wild-type cells.
越来越多的证据表明真菌中的极性生长调控与致病性相关,这引发了人们为更好地理解致病真菌中决定极化的机制而付出的巨大努力。在此,我们对植物致病担子菌玉米黑粉菌中的Spa2蛋白进行了表征,Spa2蛋白是极化体的一个著名组分,最初在酿酒酵母中被描述。玉米黑粉菌在单倍体细胞的芽殖生长和双核体的丝状生长之间表现出二态性转换。在酵母生长过程中,绿色荧光蛋白标记的Spa2蛋白以细胞周期特异性方式定位于不同的生长位点,而在菌丝生长过程中,它持续定位于菌丝尖端。与野生型细胞相比,spa2基因的缺失产生更圆的芽殖细胞和更粗的菌丝,这表明Spa2在玉米黑粉菌生长区域的确定中发挥作用。我们还研究了Spa2与肌动蛋白和微管细胞骨架之间的联系。我们发现Spa2的缺失不影响细胞骨架的组织,而且令人惊讶的是,干扰肌动蛋白丝或微管的形成并不影响Spa2的极性定位。相反,小GTP酶Rac1的缺陷似乎影响Spa2定位到顶端细胞精确位点的能力。最后,令我们惊讶的是,我们发现Spa2功能缺陷的细胞与野生型细胞一样具有致病性。