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dc.contributor.authorShrestha, Birendra Kumar
dc.contributor.authorRasmussen, Mads Skytte
dc.contributor.authorAbudu, Yakubu Princely
dc.contributor.authorBruun, Jack-Ansgar
dc.contributor.authorLarsen, Kenneth Bowitz
dc.contributor.authorAlemu, Endalkachew Ashenafi
dc.contributor.authorSjøttem, Eva
dc.contributor.authorLamark, Trond
dc.contributor.authorJohansen, Terje
dc.date.accessioned2020-04-01T07:57:11Z
dc.date.available2020-04-01T07:57:11Z
dc.date.issued2019-12-19
dc.description.abstractHuman ATG8 family proteins (ATG8s) are active in all steps of the macroautophagy pathway, and their lipidation is essential for autophagosome formation. Lipidated ATG8s anchored to the outer surface of the phagophore serve as scaffolds for binding of other core autophagy proteins and various effector proteins involved in trafficking or fusion events, whereas those at the inner surface are needed for assembly of selective autophagy substrates. Their scaffolding role depends on specific interactions between the LC3-interacting region (LIR) docking site (LDS) in ATG8s and LIR motifs in various interaction partners. LC3B is phosphorylated at Thr-50 within the LDS by serine/threonine kinase (STK) 3 and STK4. Here, we identified LIR motifs in STK3 and atypical protein kinase Cζ (PKCζ) and never in mitosis A (NIMA)-related kinase 9 (NEK9). All three kinases phosphorylated LC3B Thr-50 <i>in vitro</i>. A phospho-mimicking substitution of Thr-50 impaired binding of several LIR-containing proteins, such as ATG4B, FYVE, and coiled-coil domain-containing 1 (FYCO1), and autophagy cargo receptors p62/sequestosome 1 (SQSTM1) and neighbor of BRCA1 gene (NBR1). NEK9 knockdown or knockout enhanced degradation of the autophagy receptor and substrate p62. Of note, the suppression of p62 degradation was mediated by NEK9-mediated phosphorylation of LC3B Thr-50. Consistently, reconstitution of LC3B-KO cells with the phospho-mimicking T50E variant inhibited autophagic p62 degradation. PKCζ knockdown did not affect autophagic p62 degradation, whereas STK3/4 knockouts inhibited autophagic p62 degradation independently of LC3B Thr-50 phosphorylation. Our findings suggest that NEK9 suppresses LC3B-mediated autophagy of p62 by phosphorylating Thr-50 within the LDS of LC3B.en_US
dc.identifier.citationShrestha BK, Rasmussen MS, Abudu YP, Bruun JA, Larsen KBL, Alemu EA, Sjøttem E, Lamark T, Johansen T. NIMA-related kinase 9–mediated phosphorylation of the microtubule-associated LC3B protein at Thr-50 suppresses selective autophagy of p62/sequestosome 1 . Journal of Biological Chemistry. 2019en_US
dc.identifier.cristinIDFRIDAID 1764008
dc.identifier.doi10.1074/jbc.RA119.010068
dc.identifier.issn0021-9258
dc.identifier.issn1083-351X
dc.identifier.urihttps://hdl.handle.net/10037/17952
dc.language.isoengen_US
dc.publisherAmerican Society for Biochemistry and Molecular Biologyen_US
dc.relation.journalJournal of Biological Chemistry
dc.relation.projectIDinfo:eu-repo/grantAgreement/RCN/FRIMEDBIO/249884/Norway/Autophagy-regulated Signalosomes in Cellular Stress and Disease Pathways//en_US
dc.relation.projectIDinfo:eu-repo/grantAgreement/RCN/FRIMEDBIO/ 214448/Norway/Selective autophagy and cell signalling: Regulation of p62/SQSTM1 and functional studies of novel LIR-containing proteins//en_US
dc.rights.accessRightsopenAccessen_US
dc.rights.holderCopyright 2019 The American Society for Biochemistry and Molecular Biology, Inc.en_US
dc.subjectVDP::Mathematics and natural science: 400::Chemistry: 440::Organic chemistry: 441en_US
dc.subjectVDP::Matematikk og Naturvitenskap: 400::Kjemi: 440::Organisk kjemi: 441en_US
dc.titleNIMA-related kinase 9–mediated phosphorylation of the microtubule-associated LC3B protein at Thr-50 suppresses selective autophagy of p62/sequestosome 1en_US
dc.type.versionacceptedVersionen_US
dc.typeJournal articleen_US
dc.typeTidsskriftartikkelen_US
dc.typePeer revieweden_US


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