Transport of fungal RAB11 secretory vesicles involves myosin-5, dynein/dynactin/p25, and kinesin-1 and is independent of kinesin-3.
Transport of fungal RAB11 secretory vesicles involves myosin-5, dynein/dynactin/p25, and kinesin-1 and is independent of kinesin-3.
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DOI：
10.1091/mbc.e16-08-0566
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发表时间：
2017-04-01
影响因子：
3.3
通讯作者：
Pantazopoulou A
中科院分区：
文献类型：
--
作者：
Peñalva MA;Zhang J;Xiang X;Pantazopoulou A
在细粒曲霉中，胞外载体的分布涉及动力蛋白-1、肌球蛋白-5和动力蛋白之间的相互作用。动力蛋白复合物与这些载体的结合需要动力蛋白p25，但与早期核内体不同，它不需要Hook复合物。中性曲霉的菌丝尖端细胞是研究远距离细胞内交通的有用材料。含有RAB11同源物RabE的后高尔基分泌囊泡（SVs）参与肌球蛋白-5以及正端和负端定向微管马达，为研究微管马达和肌动蛋白马达作用于同一货物之间的相互作用提供了实验系统。通过利用f -肌动蛋白解聚释放由肌球蛋白-5聚集在顶端的SVs到微管依赖马达的事实，我们确定SVs的负端定向运输需要动力蛋白/动力蛋白超复合体。这种负端定向转运在很大程度上不受早期核内体（EEs）适应动力蛋白的Hook复合体的遗传消融的影响，但绝对需要p25的动力蛋白。因此，动力蛋白募集到两种不同的膜货物，即EEs和SVs，需要p25，强调动力蛋白尖端复合物对支架货物的重要性。最后，通过对kinesin-3和kinesin-1（分别为UncA和KinA）的零突变体和严格突变体中SVs和EEs的行为进行研究，我们证明了KinA是介导SVs顺行运输的主要激酶。因此，sv通过运动蛋白-1和肌球蛋白-5的协同顺行运输到达扁豆的顶端，并可以在动力蛋白的驱动下离开顶端。
In Aspergillus nidulans, the distribution of exocytic carriers involves interplay between kinesin-1, myosin-5, and dynein. Engagement of the dynein complex to these carriers requires dynactin p25, but, unlike that of early endosomes, it does not require the Hook complex. Hyphal tip cells of the fungus Aspergillus nidulans are useful for studying long-range intracellular traffic. Post-Golgi secretory vesicles (SVs) containing the RAB11 orthologue RabE engage myosin-5 as well as plus end– and minus end–directed microtubule motors, providing an experimental system with which to investigate the interplay between microtubule and actin motors acting on the same cargo. By exploiting the fact that depolymerization of F-actin unleashes SVs focused at the apex by myosin-5 to microtubule-dependent motors, we establish that the minus end–directed transport of SVs requires the dynein/dynactin supercomplex. This minus end–directed transport is largely unaffected by genetic ablation of the Hook complex adapting early endosomes (EEs) to dynein but absolutely requires p25 in dynactin. Thus dynein recruitment to two different membranous cargoes, namely EEs and SVs, requires p25, highlighting the importance of the dynactin pointed-end complex to scaffold cargoes. Finally, by studying the behavior of SVs and EEs in null and rigor mutants of kinesin-3 and kinesin-1 (UncA and KinA, respectively), we demonstrate that KinA is the major kinesin mediating the anterograde transport of SVs. Therefore SVs arrive at the apex of A. nidulans by anterograde transport involving cooperation of kinesin-1 with myosin-5 and can move away from the apex powered by dynein.