Extended Data Fig. 6: Unlike ACC1/Acc1, ACC2/Hfa1 is not involved in the regulation of mTORC1/TORC1 by Mal-CoA.
From: Malonyl-CoA is a conserved endogenous ATP-competitive mTORC1 inhibitor
a–c, Unlike FASN inhibition by treatment with cerulenin, exogenous overexpression of WT or a hyperactive ACC1 mutant (ACC1S79A) is not able to significantly alter Mal-CoA levels and mTORC1 activity in mammalian cells. b,c, Quantification of S6K phosphorylation (b) and Mal-K levels (c); n = 3 independent experiments. d–f, Unlike ACC1 (see Fig. 4c–e), ACC2 knockdown does not restore the increase in Mal-CoA levels or rescue the downregulation of mTORC1 caused by silencing of FASN. e, Quantification of mTORC1 activity (p-S6KT389/S6K). f, Quantification of Mal-K levels (Mal-K/loading controls); n = 3 independent experiments. g–i, C-terminal GFP tagging of the yeast ACC2 orthologue, Hfa1, does not affect TORC1 activity or cellular Mal-CoA levels. h,i, Quantification of Sch9 phosphorylation (h) and Mal-K levels (i); n = 3 independent experiments. Data in graphs are shown as the mean ± s.e.m. *P < 0.05, **P < 0.005. Source numerical data and unprocessed blots are provided.
