RT Journal Article SR Electronic T1 Threonine phosphorylation regulates the molecular assembly and signaling of EGFR in cooperation with membrane lipids JF bioRxiv FD Cold Spring Harbor Laboratory SP 2021.05.14.444132 DO 10.1101/2021.05.14.444132 A1 Ryo Maeda A1 Hiroko Tamagaki-Asahina A1 Takeshi Sato A1 Masataka Yanagawa A1 Yasushi Sako YR 2021 UL http://biorxiv.org/content/early/2021/09/06/2021.05.14.444132.abstract AB The cytoplasmic domain of the receptor tyrosine kinases (RTKs) plays roles as a phosphorylation enzyme and a protein scaffold but the regulation of these two functions is not fully understood. We here analyzed assembly of the transmembrane (TM)- juxtamembrane (JM) region of EGFR, one of the best studied species of RTKs, by combining single-pair FRET imaging and a nanodisc technique. The JM domain of EGFR contains a threonine residue that is phosphorylated after ligand association. We observed that the TM-JM peptides of EGFR form anionic lipid-induced dimers and cholesterol-induced oligomers. The two forms involve distinct molecular interactions, with a bias towards oligomer formation upon threonine phosphorylation. We further analyzed the functions of whole EGFR molecules, with or without a threonine to alanine substitution in the JM domain, in living cells. The results suggested an autoregulatory mechanism in which threonine phosphorylation of the JM domain causes a switch from kinase activation dimers to scaffolding oligomers.Competing Interest StatementThe authors have declared no competing interest.