Zheng Huawei, Guo Zhongkun, Xi Yang, Yuan Mingyue, Lin Yahong, Wu Congxian, Abubakar Yakubu Saddeeq, Dou Xianying, Li Guangpu, Wang Zonghua, Zheng Wenhui, Zhou Jie
State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, College of Life Science, Fujian Agriculture and Forestry University, Fuzhou, China.
Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, USA.
Environ Microbiol. 2017 Oct;19(10):4301-4317. doi: 10.1111/1462-2920.13896. Epub 2017 Sep 15.
Vps17 is a sorting nexin (SNX) and a component of the retromer, a protein complex mediating retrograde vesicle transport between endosomes and the trans-Golgi network. However, its role in the development and pathogenicity of filamentous fungi such as the rice blast fungus (Magnaporthe oryzae) remains unclear. We investigate the functional relationship between the SNX and the cargo-selective complex (CSC) of the fungal retromer by genetic analysis, live cell imaging and immunological assay. Our data show that the MoVps17 null mutation causes defects in growth, development and pathogenicity in M. oryzae. MoVps17 is localized to endosomes depending on the activity of phosphatidylinositol 3-kinase (PI3K), a key enzyme for fungal development and infection. Both PX and BAR domains of MoVps17 are essential for its endosomal localization and function. Furthermore, our yeast two-hybrid assays show that MoVps17 and MoVps5 can interact. Lastly, live cell imaging suggests that MoVps17 can regulate early endosome fusion and budding as well as endocytosis. Taken together, our results suggest that MoVps17 specifically functions as a retromer component with CSC and also plays a distinct role in the regulation of endosome dynamics during fungal development and plant infection.
Vps17是一种分选连接蛋白(SNX),也是回收转运复合体的一个组成部分,该蛋白复合体介导内体与反式高尔基体网络之间的逆行囊泡运输。然而,其在丝状真菌(如稻瘟病菌,Magnaporthe oryzae)的发育和致病性中的作用仍不清楚。我们通过遗传分析、活细胞成像和免疫测定来研究真菌回收转运复合体的SNX与货物选择复合体(CSC)之间的功能关系。我们的数据表明,MoVps17基因敲除突变会导致稻瘟病菌在生长、发育和致病性方面出现缺陷。MoVps17定位于内体,这取决于磷脂酰肌醇3激酶(PI3K)的活性,PI3K是真菌发育和感染的关键酶。MoVps17的PX和BAR结构域对其在内体的定位和功能都至关重要。此外,我们的酵母双杂交试验表明MoVps17和MoVps5可以相互作用。最后,活细胞成像表明MoVps17可以调节早期内体融合、出芽以及内吞作用。综上所述,我们的结果表明MoVps17作为回收转运复合体的一个组成部分,与CSC具有特定功能,并且在真菌发育和植物感染过程中对内体动态的调节中也发挥着独特作用。