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. Author manuscript; available in PMC: 2011 Apr 23.
Published in final edited form as: Mol Cell. 2010 Apr 23;38(2):250–264. doi: 10.1016/j.molcel.2010.02.033

Figure 7.

Figure 7

Rim15 is a downstream target of Sic1.

(A) Deletion of RIM15 or GCN4, but not PHO4, suppresses the upregulation of autophagy in sic1Δ cells. Wild-type (TN124), atg1Δ (HAY572), sic1Δ (ZFY098), pho4Δ (ZFY135), pho4Δ sic1Δ (ZFY141), rim15Δ (ZFY100), rim15Δ sic1Δ (ZFY116), gcn4Δ (ZFY111) and gcn4Δ sic1Δ (ZFY115) cells were analyzed by the Pho8Δ60 assay, as described in Figure 2D.

(B) and (C) Overexpression of Sic1 inhibits autophagy in wild-type, pho4Δ and gcn4Δ cells, but not rim15Δ cells. (B) Wild type (W303-1B), rim15Δ (ZFY132), pho4Δ (ZFY170), gcn4Δ (ZFY131), GAL1-SIC1 (ZFY184), rim15Δ GAL1-SIC1 (ZFY187), pho4Δ GAL1-SIC1 (ZFY188) and gcn4Δ GAL1-SIC1 (ZFY185) cells were analyzed by the GFP-Atg8 processing assay, as described in Figure 2A. (C) Wild-type (ZFY202), rim15Δ (ZFY204), pho4Δ (ZFY252), gcn4Δ (ZFY251), GAL1-SIC1 (ZFY203), rim15Δ GAL1-SIC1 (ZFY249), pho4Δ GAL1-SIC1 (ZFY250) and gcn4Δ GAL1-SIC1 (ZFY248) cells were analyzed by the Pho8Δ60 assay, as described in Figure 2B. The difference of the Pho8Δ60 activity between the absence and presence of Sic overexpression was normalized to the difference in wild-type cells treated with nitrogen starvation, which was set to 100%. Error bars indicate the SD of three independent experiments.

(D) and (E) Overexpression of Rim15 partially suppresses the inhibitory effect of Sic1 overexpression on autophagy. (D) Wild-type (W303-1B), GAL1-RIM15 (ZFY192), GAL1-SIC1 (ZFY184) and GAL1-SIC1 GAL1-RIM15 (ZFY193) cells were analyzed by the GFP-Atg8 processing assay, as described in Figure 2A. (E) Wild-type (ZFY202), GAL1-RIM15 (ZFY253), GAL1-SIC1 (ZFY203) and GAL1-SIC1 GAL1-RIM15 (ZFY254) cells were analyzed by the Pho8Δ60 assay, as described in Figure 2B.

(F) Deletion of RIM15 blocks the induction of autophagy upon overexpression of Pho85 in pcl5Δ pho80Δ cells. Wild-type (W303-1B), pcl5Δ pho80Δ (ZFY207), GAL1-PHO85 (ZFY209), pcl5Δ pho80Δ GAL1-PHO85 (ZFY208) and pcl5Δ pho80Δ rim15Δ GAL1-PHO85 (ZFY216) cells were analyzed by the GFP-Atg8 processing assay, as described in Figure 4A. See also Figure S7.