Schuchardt Isabel, Assmann Daniela, Thines Eckhard, Schuberth Christian, Steinberg Gero
Max-Planck-Institut für Terrestrische Mikrobiologie, Karl-von-Frisch-Strasse, D-35043 Marburg, Germany.
Mol Biol Cell. 2005 Nov;16(11):5191-201. doi: 10.1091/mbc.e05-04-0272. Epub 2005 Aug 24.
Long-distance transport is crucial for polar-growing cells, such as neurons and fungal hyphae. Kinesins and myosins participate in this process, but their functional interplay is poorly understood. Here, we investigate the role of kinesin motors in hyphal growth of the plant pathogen Ustilago maydis. Although the microtubule plus-ends are directed to the hyphal tip, of all 10 kinesins analyzed, only conventional kinesin (Kinesin-1) and Unc104/Kif1A-like kinesin (Kinesin-3) were up-regulated in hyphae and they are essential for extended hyphal growth. deltakin1 and deltakin3 mutant hyphae grew irregular and remained short, but they were still able to grow polarized. No additional phenotype was detected in deltakin1rkin3 double mutants, but polarity was lost in deltamyo5rkin1 and deltamyo5rkin3 mutant cells, suggesting that kinesins and class V myosin cooperate in hyphal growth. Consistent with such a role in secretion, fusion proteins of green fluorescent protein and Kinesin-1, Myosin-V, and Kinesin-3 accumulate in the apex of hyphae, a region where secretory vesicles cluster to form the fungal Spitzenkörper. Quantitative assays revealed a role of Kin3 in secretion of acid phosphatase, whereas Kin1 was not involved. Our data demonstrate that just two kinesins and at least one myosin support hyphal growth.
长距离运输对于极性生长的细胞至关重要,比如神经元和真菌菌丝。驱动蛋白和肌球蛋白参与这一过程,但其功能相互作用却鲜为人知。在此,我们研究了驱动蛋白在植物病原体玉米黑粉菌菌丝生长中的作用。尽管微管正端指向菌丝尖端,但在分析的所有10种驱动蛋白中,只有传统驱动蛋白(驱动蛋白-1)和Unc104/Kif1A样驱动蛋白(驱动蛋白-3)在菌丝中上调,且它们对于菌丝的延伸生长至关重要。缺失Kin1和缺失Kin3的突变菌丝生长不规则且短小,但仍能极性生长。在缺失Kin1缺失Kin3的双突变体中未检测到其他表型,但在缺失Myo5缺失Kin1和缺失Myo5缺失Kin3的突变细胞中极性丧失,这表明驱动蛋白和V类肌球蛋白在菌丝生长中协同作用。与在分泌中的这种作用一致,绿色荧光蛋白与驱动蛋白-1、肌球蛋白-V和驱动蛋白-3的融合蛋白聚集在菌丝顶端,分泌囊泡在此聚集形成真菌的顶体。定量分析揭示了Kin3在酸性磷酸酶分泌中的作用,而Kin1不参与此过程。我们的数据表明,仅两种驱动蛋白和至少一种肌球蛋白支持菌丝生长。