Seiler Stephan, Plamann Michael
Institut für Mikrobiologie und Genetik, Abteilung Molekulare Mikrobiologie, Universität Göttingen, D-37077 Göttingen, Germany.
Mol Biol Cell. 2003 Nov;14(11):4352-64. doi: 10.1091/mbc.e02-07-0433. Epub 2003 Sep 5.
Cellular polarity is a fundamental property of every cell. Due to their extremely fast growth rate (>/=1 microm/s) and their highly elongated form, filamentous fungi represent a prime example of polarized growth and are an attractive model for the analysis of fundamental mechanisms underlying cellular polarity. To identify the critical components that contribute to polarized growth, we developed a large-scale genetic screen for the isolation of conditional mutants defective in this process in the model fungus Neurospora crassa. Phenotypic analysis and complementation tests of ca. 950 mutants identified more than 100 complementation groups that define 21 distinct morphological classes. The phenotypes include polarity defects over the whole hypha, more specific defects localized to hyphal tips or subapical regions, and defects in branch formation and growth directionality. To begin converting this mutant collection into meaningful biological information, we identified the defective genes in 45 mutants covering all phenotypic classes. These genes encode novel proteins as well as proteins which 1) regulate the actin or microtubule cytoskeleton, 2) are kinases or components of signal transduction pathways, 3) are part of the secretory pathway, or 4) have functions in cell wall formation or membrane biosynthesis. These findings highlight the dynamic nature of a fungal hypha and establish a molecular model for studies of hyphal growth and polarity.
细胞极性是每个细胞的基本特性。丝状真菌由于其极快的生长速度(≥1微米/秒)和高度细长的形态,成为极性生长的典型例子,是分析细胞极性潜在基本机制的有吸引力的模型。为了鉴定对极性生长有贡献的关键成分,我们开发了一种大规模遗传筛选方法,用于在模式真菌粗糙脉孢菌中分离在此过程中有缺陷的条件突变体。对约950个突变体的表型分析和互补试验确定了100多个互补组,这些互补组定义了21个不同的形态学类别。这些表型包括整个菌丝上的极性缺陷、更特定的局限于菌丝顶端或亚顶端区域的缺陷,以及分支形成和生长方向性方面的缺陷。为了开始将这个突变体库转化为有意义的生物学信息,我们鉴定了涵盖所有表型类别的45个突变体中的缺陷基因。这些基因编码新蛋白以及以下几类蛋白:1)调节肌动蛋白或微管细胞骨架的蛋白,2)激酶或信号转导途径的成分,3)分泌途径的一部分,或4)在细胞壁形成或膜生物合成中起作用的蛋白。这些发现突出了真菌菌丝的动态性质,并建立了一个用于研究菌丝生长和极性的分子模型。