Supplementary Figure 9: Phosphorylation of TOR-Dependent Sites S614 and S617 Mediates Decapping and Degradation of ATG8 Transcripts, Related to Fig. 4.
From: A conserved mechanism of TOR-dependent RCK-mediated mRNA degradation regulates autophagy
(a) Yeast cells under the indicated conditions expressing either WT Dcp2 or an equivalent protein containing either an S > D mutation or S > A mutation at positions S614 and S617 were assayed for decapping of ATG8 transcripts over 30 min by a PCR assay and visualized by ethidium bromide as in Fig. 3b. (b) Decapping assay: Densitometry of n = 3 independent assays Dcp2 containing an S > D mutation or S > A mutation at a single position S614 or S617 were assayed for degradation as in Fig. 4b. Degradation (c) and Decapping (d) assays conducted on yeast cells expressing WT or Dcp2 with single mutations; decapping assay conducted as in Supplementary Fig. 3a and B. All experiments the results of n = 3 independent assays + / − SD. Student’s t-test, ∗p < 0.05,∗∗p < 0.01,∗∗∗p < 0.001,∗∗∗∗p < 0.0001.
