Konzack Sven, Rischitor Patricia E, Enke Cathrin, Fischer Reinhard
Department of Microbiology, University of Marburg, Max-Planck-Institute for Terrestrial Microbiology, D-35043 Marburg, Germany.
Mol Biol Cell. 2005 Feb;16(2):497-506. doi: 10.1091/mbc.e04-02-0083. Epub 2004 Nov 24.
Polarized growth in filamentous fungi requires the integrity of the microtubule (MT) cytoskeleton. We found that growing MTs in Aspergillus nidulans merge at the center of fast growing tips and discovered that a kinesin motor protein, KipA, related to Tea2p of Schizosaccharomyces pombe, is required for this process. In a DeltakipA strain, MT plus ends reach the tip but show continuous lateral movement. Hyphae lose directionality and grow in curves, apparently due to mislocalization of the vesicle supply center (Spitzenkörper) in the apex. Green fluorescent protein (GFP)-KipA accumulates at MT plus ends, whereas a KipA rigor mutant protein, GFP-KipA(G223E), coated MTs evenly. These findings suggest that KipA requires its intrinsic motor activity to reach the MT plus end. Using KipA as an MT plus-end marker, we found bidirectional organization of MTs and determined the locations of microtubule organizing centers at nuclei, in the cytoplasm, and at septa.
丝状真菌中的极性生长需要微管(MT)细胞骨架的完整性。我们发现,构巢曲霉中生长的微管在快速生长的顶端中心合并,并发现一种驱动蛋白KipA(与粟酒裂殖酵母的Tea2p相关)参与了这一过程。在缺失KipA的菌株中,微管正端到达顶端,但显示出持续的横向运动。菌丝失去方向性并弯曲生长,这显然是由于顶端的囊泡供应中心(Spitzenkörper)定位错误所致。绿色荧光蛋白(GFP)标记的KipA在微管正端积累,而KipA的僵直突变蛋白GFP-KipA(G223E)则均匀地覆盖在微管上。这些发现表明,KipA需要其内在的驱动活性才能到达微管正端。以KipA作为微管正端标记,我们发现了微管的双向组织,并确定了细胞核、细胞质和隔膜处微管组织中心的位置。