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Vps9 Family Protein Muk1 Is the Second Rab5 Guanosine Nucleotide Exchange Factor in Budding Yeast

机译:Vps9家族蛋白Muk1是芽酵母中的第二个Rab5鸟苷核苷酸交换因子。

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摘要

VPS9 domains can act as guanosine nucleotide exchange factors (GEFs) against small G proteins of the Rab5 family. Saccharomyces cerevisiae vps9Δ mutants have trafficking defects considerably less severe than multiple deletions of the three cognate Rab5 paralogs (Vps21, Ypt52, and Ypt53). Here, we show that Muk1, which also contains a VPS9 domain, acts as a second GEF against Vps21, Ypt52, and Ypt53. Muk1 is partially redundant with Vps9 in vivo, with vps9Δ muk1Δ double mutant cells displaying hypersensitivity to temperature and ionic stress, as well as profound impairments in endocytic and Golgi endosome trafficking, including defects in sorting through the multivesicular body. Cells lacking both Vps9 and Muk1 closely phenocopy double and triple knock-out strains lacking Rab5 paralogs. Microscopy and overexpression experiments demonstrate that Vps9 and Muk1 have distinct localization determinants. These experiments establish Muk1 as the second Rab5 GEF in budding yeast.
机译:VPS9域可以充当针对Rab5家族小G蛋白的鸟嘌呤核苷酸交换因子(GEF)。酿酒酵母vps9Δ突变体具有比三个同源Rab5旁系同源物(Vps21,Ypt52和Ypt53)的多次缺失严重得多的运输缺陷。在这里,我们显示Muk1(也包含VPS9域)充当针对Vps21,Ypt52和Ypt53的第二个GEF。 Muk1在体内与Vps9部分重叠,vps9Δmuk1Δ双突变细胞对温度和离子应力表现出超敏性,并且在胞吞和高尔基体内体运输中受到严重损害,包括通过多囊体分选的缺陷。缺乏Vps9和Muk1的细胞在表型上缺乏Rab5旁系同源物的双重和三重敲除菌株。显微镜和过表达实验表明,Vps9和Muk1具有不同的定位决定因素。这些实验将Muk1确定为发芽酵母中的第二个Rab5 GEF。

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