PMID- 30827897 OWN - NLM STAT- MEDLINE DCOM- 20191107 LR - 20211204 IS - 1878-1551 (Electronic) IS - 1534-5807 (Print) IS - 1534-5807 (Linking) VI - 49 IP - 1 DP - 2019 Apr 8 TI - Phosphorylation of Syntaxin 17 by TBK1 Controls Autophagy Initiation. PG - 130-144.e6 LID - S1534-5807(19)30055-3 [pii] LID - 10.1016/j.devcel.2019.01.027 [doi] AB - Syntaxin 17 (Stx17) has been implicated in autophagosome-lysosome fusion. Here, we report that Stx17 functions in assembly of protein complexes during autophagy initiation. Stx17 is phosphorylated by TBK1 whereby phospho-Stx17 controls the formation of the ATG13(+)FIP200(+) mammalian pre-autophagosomal structure (mPAS) in response to induction of autophagy. TBK1 phosphorylates Stx17 at S202. During autophagy induction, Stx17(pS202) transfers from the Golgi, where its steady-state pools localize, to the ATG13(+)FIP200(+) mPAS. Stx17(pS202) was in complexes with ATG13 and FIP200, whereas its non-phosphorylatable mutant Stx17(S202A) was not. Stx17 or TBK1 knockouts blocked ATG13 and FIP200 puncta formation. Stx17 or TBK1 knockouts reduced the formation of ATG13 protein complexes with FIP200 and ULK1. Endogenous Stx17(pS202) colocalized with LC3B following induction of autophagy. Stx17 knockout diminished LC3 response and reduced sequestration of the prototypical bulk autophagy cargo lactate dehydrogenase. We conclude that Stx17 is a TBK1 substrate and that together they orchestrate assembly of mPAS. CI - Published by Elsevier Inc. FAU - Kumar, Suresh AU - Kumar S AD - Autophagy Inflammation and Metabolism Center of Biomedical Research Excellence, University of New Mexico Health Sciences Center, Albuquerque, NM 87131, USA; Department of Molecular Genetics and Microbiology, University of New Mexico Health Sciences Center, Albuquerque, NM 87131, USA. FAU - Gu, Yuexi AU - Gu Y AD - Autophagy Inflammation and Metabolism Center of Biomedical Research Excellence, University of New Mexico Health Sciences Center, Albuquerque, NM 87131, USA; Department of Molecular Genetics and Microbiology, University of New Mexico Health Sciences Center, Albuquerque, NM 87131, USA. FAU - Abudu, Yakubu Princely AU - Abudu YP AD - Molecular Cancer Research Group, Institute of Medical Biology, University of Tromso, The Arctic University of Norway, Tromso 9037, Norway. FAU - Bruun, Jack-Ansgar AU - Bruun JA AD - Molecular Cancer Research Group, Institute of Medical Biology, University of Tromso, The Arctic University of Norway, Tromso 9037, Norway. FAU - Jain, Ashish AU - Jain A AD - Department of Molecular Cell Biology, Centre for Cancer Biomedicine, University of Oslo and Institute for Cancer Research, The Norwegian Radium Hospital, Oslo 0379, Norway. FAU - Farzam, Farzin AU - Farzam F AD - Departments of Physics and Astronomy, University of New Mexico, Albuquerque, NM 87131, USA. FAU - Mudd, Michal AU - Mudd M AD - Autophagy Inflammation and Metabolism Center of Biomedical Research Excellence, University of New Mexico Health Sciences Center, Albuquerque, NM 87131, USA; Department of Molecular Genetics and Microbiology, University of New Mexico Health Sciences Center, Albuquerque, NM 87131, USA. FAU - Anonsen, Jan Haug AU - Anonsen JH AD - Department of Biosciences IBV Mass Spectrometry and Proteomics Unit, University of Oslo, Oslo 0371, Norway. FAU - Rusten, Tor Erik AU - Rusten TE AD - Department of Molecular Cell Biology, Centre for Cancer Biomedicine, University of Oslo and Institute for Cancer Research, The Norwegian Radium Hospital, Oslo 0379, Norway. FAU - Kasof, Gary AU - Kasof G AD - Cell Signaling Technology, Danvers, MA 01923, USA. FAU - Ktistakis, Nicholas AU - Ktistakis N AD - Signalling Programme, The Babraham Institute, Cambridge CB22 3AT, UK. FAU - Lidke, Keith A AU - Lidke KA AD - Departments of Physics and Astronomy, University of New Mexico, Albuquerque, NM 87131, USA. FAU - Johansen, Terje AU - Johansen T AD - Molecular Cancer Research Group, Institute of Medical Biology, University of Tromso, The Arctic University of Norway, Tromso 9037, Norway. FAU - Deretic, Vojo AU - Deretic V AD - Autophagy Inflammation and Metabolism Center of Biomedical Research Excellence, University of New Mexico Health Sciences Center, Albuquerque, NM 87131, USA; Department of Molecular Genetics and Microbiology, University of New Mexico Health Sciences Center, Albuquerque, NM 87131, USA. Electronic address: vderetic@salud.unm.edu. LA - eng GR - BBS/E/B/000C0413/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom GR - R01 AI042999/AI/NIAID NIH HHS/United States GR - BB/K019155/1/BB_/Biotechnology and Biological Sciences Research Council/United Kingdom GR - P20 GM121176/GM/NIGMS NIH HHS/United States GR - R37 AI042999/AI/NIAID NIH HHS/United States GR - R01 AI111935/AI/NIAID NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20190228 PL - United States TA - Dev Cell JT - Developmental cell JID - 101120028 RN - 0 (ATG13 protein, human) RN - 0 (Adaptor Proteins, Signal Transducing) RN - 0 (Autophagy-Related Proteins) RN - 0 (Multiprotein Complexes) RN - 0 (Qa-SNARE Proteins) RN - 0 (RB1CC1 protein, human) RN - 0 (STX17 protein, human) RN - EC 2.7.10.1 (Protein-Tyrosine Kinases) RN - EC 2.7.11.1 (Protein Serine-Threonine Kinases) RN - EC 2.7.11.1 (TBK1 protein, human) SB - IM MH - Adaptor Proteins, Signal Transducing/genetics MH - Autophagosomes/metabolism MH - Autophagy/*genetics MH - Autophagy-Related Proteins/genetics MH - Gene Knockout Techniques MH - Golgi Apparatus/genetics/metabolism MH - HEK293 Cells MH - HeLa Cells MH - Humans MH - Lysosomes/metabolism MH - Membrane Fusion/genetics MH - Multiprotein Complexes/*genetics/metabolism MH - Mutation/genetics MH - Phosphorylation MH - Protein Serine-Threonine Kinases/*genetics MH - Protein-Tyrosine Kinases/genetics MH - Qa-SNARE Proteins/*genetics MH - Signal Transduction/genetics PMC - PMC6907693 MID - NIHMS1520655 OTO - NOTNLM OT - TBK1 OT - ULK1 OT - autophagy OT - pre-autophagosomal structure COIS- Declaration of Interest: Gary Kasof is employed by Cell Signaling Technology. EDAT- 2019/03/05 06:00 MHDA- 2019/11/08 06:00 PMCR- 2020/04/08 CRDT- 2019/03/05 06:00 PHST- 2018/10/12 00:00 [received] PHST- 2018/12/16 00:00 [revised] PHST- 2019/01/30 00:00 [accepted] PHST- 2019/03/05 06:00 [pubmed] PHST- 2019/11/08 06:00 [medline] PHST- 2019/03/05 06:00 [entrez] PHST- 2020/04/08 00:00 [pmc-release] AID - S1534-5807(19)30055-3 [pii] AID - 10.1016/j.devcel.2019.01.027 [doi] PST - ppublish SO - Dev Cell. 2019 Apr 8;49(1):130-144.e6. doi: 10.1016/j.devcel.2019.01.027. Epub 2019 Feb 28.