Yang Shu, Rosenwald Anne
Department of Biology, Georgetown University, Washington, DC 20057.
Department of Biology, Georgetown University, Washington, DC 20057
G3 (Bethesda). 2017 Feb 9;7(2):333-341. doi: 10.1534/g3.116.035998.
In , Arl1 and Ypt6, two small GTP-binding proteins that regulate membrane traffic in the secretory and endocytic pathways, are also necessary for autophagy. To gain information about potential partners of Arl1 and Ypt6 specifically in autophagy, we carried out a high copy number suppressor screen to identify genes that when overexpressed suppress the rapamycin sensitivity phenotype of Δ and Δ strains at 37°. From the screen results, we selected , , , , , , and , either membrane traffic or autophagy regulators, to further test whether they can suppress the specific autophagy defects of Δ and Δ strains. As a result, we identified , , and to be specific suppressors for the Δ strain, and and for the Δ strain. Through this screen, we were able to confirm several membrane traffic and autophagy regulators that have novel relationships with Arl1 and Ypt6 during autophagy.
在酵母中,Arl1和Ypt6这两种调节分泌和内吞途径中膜运输的小GTP结合蛋白对于自噬也是必需的。为了获得有关Arl1和Ypt6在自噬中潜在伙伴的信息,我们进行了高拷贝数抑制子筛选,以鉴定那些在过表达时能抑制Δarl1和Δypt6菌株在37°时对雷帕霉素敏感性表型的基因。从筛选结果中,我们选择了VPS39、VPS41、VPS42、VPS45、HOPS复合体亚基Vps33、Ypt1和Sec15,它们要么是膜运输调节因子,要么是自噬调节因子,以进一步测试它们是否能抑制Δarl1和Δypt6菌株的特定自噬缺陷。结果,我们鉴定出VPS39、VPS41和VPS42是Δarl1菌株的特异性抑制子,而VPS45和Sec15是Δypt6菌株的特异性抑制子。通过这个筛选,我们能够确认几种在自噬过程中与Arl1和Ypt6具有新关系的膜运输和自噬调节因子。