A High Copy Suppressor Screen for Autophagy Defects in Saccharomyces arl1Δ and ypt6Δ Strains

摘要

In Saccharomyces cerevisiae , [Arl1][1] and [Ypt6][2], 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][1] and [Ypt6][2] specifically in autophagy, we carried out a high copy number suppressor screen to identify genes that when overexpressed suppress the rapamycin sensitivity phenotype of [arl1][1] Δ and [ypt6][2] Δ strains at 37°. From the screen results, we selected [COG4][3] , [SNX4][4] , [TAX4][5] , [IVY1][6] , [PEP3][7] , [SLT2][8] , and [ATG5][9] , either membrane traffic or autophagy regulators, to further test whether they can suppress the specific autophagy defects of [arl1][1] Δ and [ypt6][2] Δ strains. As a result, we identified [COG4][3] , [SNX4][4] , and [TAX4][5] to be specific suppressors for the [arl1][1] Δ strain, and [IVY1][6] and [ATG5][9] for the [ypt6][2] Δ strain. Through this screen, we were able to confirm several membrane traffic and autophagy regulators that have novel relationships with [Arl1][1] and [Ypt6][2] during autophagy. [1]: http://www.yeastgenome.org/locus/S000000368/overview [2]: http://www.yeastgenome.org/locus/S000004252/overview [3]: http://www.yeastgenome.org/locus/S000006309/overview [4]: http://www.yeastgenome.org/locus/S000003573/overview [5]: http://www.yeastgenome.org/locus/S000003619/overview [6]: http://www.yeastgenome.org/locus/S000002637/overview [7]: http://www.yeastgenome.org/locus/S000004138/overview [8]: http://www.yeastgenome.org/locus/S000001072/overview [9]: http://www.yeastgenome.org/locus/S000006070/overview