Bakula, DanielaDanielaBakulaMüller, Amelie JAmelie JMüllerZuleger, TheresiaTheresiaZulegerTakacs, ZsuzsannaZsuzsannaTakacsFranz-Wachtel, MiritaMiritaFranz-WachtelThost, Ann-KatrinAnn-KatrinThostBrigger, DanielDanielBriggerTschan, MarioMarioTschan0000-0001-5897-3647Frickey, TancredTancredFrickeyRobenek, HorstHorstRobenekMacek, BorisBorisMacekProikas-Cezanne, TassulaTassulaProikas-Cezanne2024-10-252024-10-252017-05-31https://boris-portal.unibe.ch/handle/20.500.12422/154412Autophagy is controlled by AMPK and mTOR, both of which associate with ULK1 and control the production of phosphatidylinositol 3-phosphate (PtdIns3P), a prerequisite for autophagosome formation. Here we report that WIPI3 and WIPI4 scaffold the signal control of autophagy upstream of PtdIns3P production and have a role in the PtdIns3P effector function of WIPI1-WIPI2 at nascent autophagosomes. In response to LKB1-mediated AMPK stimulation, WIPI4-ATG2 is released from a WIPI4-ATG2/AMPK-ULK1 complex and translocates to nascent autophagosomes, controlling their size, to which WIPI3, in complex with FIP200, also contributes. Upstream, WIPI3 associates with AMPK-activated TSC complex at lysosomes, regulating mTOR. Our WIPI interactome analysis reveals the scaffold functions of WIPI proteins interconnecting autophagy signal control and autophagosome formation. Our functional kinase screen uncovers a novel regulatory link between LKB1-mediated AMPK stimulation that produces a direct signal via WIPI4, and we show that the AMPK-related kinases NUAK2 and BRSK2 regulate autophagy through WIPI4.en500 - Science::570 - Life sciences; biology600 - Technology::610 - Medicine & healtharticle10.7892/boris.1053002856106610.1038/ncomms15637