Fig. 1: Identification and analysis of PA binding mTOR.

a Screening of PA binding proteins in vitro and test mTOR interacting with PA in vivo. Experimental setup: cell lysates of 5-d-old mycelia were incubated with PA beads. The bound proteins were subsequently eluted, separated by SDS-PAGE, and analyzed by liquid chromatography coupled to high-resolution tandem mass spectrum (LC-MS/MS). An aliquot of the cell lysates were used to isolate the PA-mTOR complex by using an anti-mTOR antibody to immunoprecipitate mTOR. The bound lipids were extracted and analyzed by LC-MS/MS. b Scatter plot of abundance-versus-sequence length of identified PA binding proteins. Arrows mark examples of PA binding mTOR. c Features of mTOR protein in G. lingzhi. Conserve domain analysis of G. lingzhi mTOR was calculated online by NCBI Conserved Domains Database. d Characterization of the PA binding domain of mTOR. Pulldown of GST or the recombinant, GST-tagged, indicated regions of mTOR interaction with the PA beads (Top panel). An aliquot of the protein samples (starting material, input) added to the PA beads was analysed in parallel (bottom panel). PA-binding was detected by immunoblotting using a GST-specific antibody. e Schematic of serial deletions of the GST-fusion proteins of the mTOR used in (d) and the relative strength of binding to PA. FL, full-length mTOR (amino acids 1 to 2388); A, amino acids 1 to 1100; B, amino acids 1101 to 2388; C, amino acids 1101 to 2284; D, amino acids 1208 to 1992; E, amino acids 1101 to 1857; F, amino acids 1859 to 1965. f The FRB sequences alignment were performed using clustal omega. Accession numbers for the aligned proteins were indicated in Supplementary Fig. 1.