Schuster M, Kilaru S, Latz M, Steinberg G
Biosciences, University of Exeter, Exeter EX4 4QD, UK.
Biosciences, University of Exeter, Exeter EX4 4QD, UK.
Fungal Genet Biol. 2015 Jun;79:141-9. doi: 10.1016/j.fgb.2015.03.005. Epub 2015 Apr 7.
The microtubule cytoskeleton supports vital processes in fungal cells, including hyphal growth and mitosis. Consequently, it is a target for fungicides, such as benomyl. The use of fluorescent fusion proteins to illuminate microtubules and microtubule-associated proteins has led to a break-through in our understanding of their dynamics and function in fungal cells. Here, we introduce fluorescent markers to visualize microtubules and accessory proteins in the wheat pathogen Zymoseptoria tritici. We fused enhanced green-fluorescent protein to α-tubulin (ZtTub2), to ZtPeb1, a homologue of the mammalian plus-end binding protein EB1, and to ZtGrc1, a component of the minus-end located γ-tubulin ring complex, involved in the nucleation of microtubules. In vivo observation confirms the localization and dynamic behaviour of all three markers. These marker proteins are useful tools for understanding the organization and importance of the microtubule cytoskeleton in Z. tritici.
微管细胞骨架支持真菌细胞中的重要过程,包括菌丝生长和有丝分裂。因此,它是杀菌剂(如苯菌灵)的作用靶点。使用荧光融合蛋白来照亮微管和微管相关蛋白,使我们对它们在真菌细胞中的动态和功能的理解有了突破。在这里,我们引入荧光标记来可视化小麦病原体小麦黄斑叶枯病菌中的微管和辅助蛋白。我们将增强型绿色荧光蛋白与α-微管蛋白(ZtTub2)、与哺乳动物正端结合蛋白EB1的同源物ZtPeb1以及与位于负端的γ-微管蛋白环复合物的一个组分ZtGrc1融合,γ-微管蛋白环复合物参与微管的成核。体内观察证实了所有三种标记物的定位和动态行为。这些标记蛋白是理解小麦黄斑叶枯病菌中微管细胞骨架的组织和重要性的有用工具。