Phosphorylation of the mitochondrial autophagy receptor Nix enhances its interaction with LC3 proteins

Abstract The mitophagy receptor Nix interacts with LC3/GABARAP proteins, targeting mitochondria into autophagosomes for degradation. Here we present evidence for phosphorylation-driven regulation of the Nix:LC3B interaction. Isothermal titration calorimetry and NMR indicate a ~100 fold enhanced affi...

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Autores principales:	Vladimir V. Rogov, Hironori Suzuki, Mija Marinković, Verena Lang, Ryuichi Kato, Masato Kawasaki, Maja Buljubašić, Matilda Šprung, Natalia Rogova, Soichi Wakatsuki, Anne Hamacher-Brady, Volker Dötsch, Ivan Dikic, Nathan R. Brady, Ivana Novak
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/505fa4ea976e48f19f058f1285520910
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spelling	oai:doaj.org-article:505fa4ea976e48f19f058f12855209102021-12-02T16:06:45ZPhosphorylation of the mitochondrial autophagy receptor Nix enhances its interaction with LC3 proteins10.1038/s41598-017-01258-62045-2322https://doaj.org/article/505fa4ea976e48f19f058f12855209102017-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-01258-6https://doaj.org/toc/2045-2322Abstract The mitophagy receptor Nix interacts with LC3/GABARAP proteins, targeting mitochondria into autophagosomes for degradation. Here we present evidence for phosphorylation-driven regulation of the Nix:LC3B interaction. Isothermal titration calorimetry and NMR indicate a ~100 fold enhanced affinity of the serine 34/35-phosphorylated Nix LC3-interacting region (LIR) to LC3B and formation of a very rigid complex compared to the non-phosphorylated sequence. Moreover, the crystal structure of LC3B in complex with the Nix LIR peptide containing glutamic acids as phosphomimetic residues and NMR experiments revealed that LIR phosphorylation stabilizes the Nix:LC3B complex via formation of two additional hydrogen bonds between phosphorylated serines of Nix LIR and Arg11, Lys49 and Lys51 in LC3B. Substitution of Lys51 to Ala in LC3B abrogates binding of a phosphomimetic Nix mutant. Functionally, serine 34/35 phosphorylation enhances autophagosome recruitment to mitochondria in HeLa cells. Together, this study provides cellular, biochemical and biophysical evidence that phosphorylation of the LIR domain of Nix enhances mitophagy receptor engagement.Vladimir V. RogovHironori SuzukiMija MarinkovićVerena LangRyuichi KatoMasato KawasakiMaja BuljubašićMatilda ŠprungNatalia RogovaSoichi WakatsukiAnne Hamacher-BradyVolker DötschIvan DikicNathan R. BradyIvana NovakNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-12 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Vladimir V. Rogov Hironori Suzuki Mija Marinković Verena Lang Ryuichi Kato Masato Kawasaki Maja Buljubašić Matilda Šprung Natalia Rogova Soichi Wakatsuki Anne Hamacher-Brady Volker Dötsch Ivan Dikic Nathan R. Brady Ivana Novak Phosphorylation of the mitochondrial autophagy receptor Nix enhances its interaction with LC3 proteins
description	Abstract The mitophagy receptor Nix interacts with LC3/GABARAP proteins, targeting mitochondria into autophagosomes for degradation. Here we present evidence for phosphorylation-driven regulation of the Nix:LC3B interaction. Isothermal titration calorimetry and NMR indicate a ~100 fold enhanced affinity of the serine 34/35-phosphorylated Nix LC3-interacting region (LIR) to LC3B and formation of a very rigid complex compared to the non-phosphorylated sequence. Moreover, the crystal structure of LC3B in complex with the Nix LIR peptide containing glutamic acids as phosphomimetic residues and NMR experiments revealed that LIR phosphorylation stabilizes the Nix:LC3B complex via formation of two additional hydrogen bonds between phosphorylated serines of Nix LIR and Arg11, Lys49 and Lys51 in LC3B. Substitution of Lys51 to Ala in LC3B abrogates binding of a phosphomimetic Nix mutant. Functionally, serine 34/35 phosphorylation enhances autophagosome recruitment to mitochondria in HeLa cells. Together, this study provides cellular, biochemical and biophysical evidence that phosphorylation of the LIR domain of Nix enhances mitophagy receptor engagement.
format	article
author	Vladimir V. Rogov Hironori Suzuki Mija Marinković Verena Lang Ryuichi Kato Masato Kawasaki Maja Buljubašić Matilda Šprung Natalia Rogova Soichi Wakatsuki Anne Hamacher-Brady Volker Dötsch Ivan Dikic Nathan R. Brady Ivana Novak
author_facet	Vladimir V. Rogov Hironori Suzuki Mija Marinković Verena Lang Ryuichi Kato Masato Kawasaki Maja Buljubašić Matilda Šprung Natalia Rogova Soichi Wakatsuki Anne Hamacher-Brady Volker Dötsch Ivan Dikic Nathan R. Brady Ivana Novak
author_sort	Vladimir V. Rogov
title	Phosphorylation of the mitochondrial autophagy receptor Nix enhances its interaction with LC3 proteins
title_short	Phosphorylation of the mitochondrial autophagy receptor Nix enhances its interaction with LC3 proteins
title_full	Phosphorylation of the mitochondrial autophagy receptor Nix enhances its interaction with LC3 proteins
title_fullStr	Phosphorylation of the mitochondrial autophagy receptor Nix enhances its interaction with LC3 proteins
title_full_unstemmed	Phosphorylation of the mitochondrial autophagy receptor Nix enhances its interaction with LC3 proteins
title_sort	phosphorylation of the mitochondrial autophagy receptor nix enhances its interaction with lc3 proteins
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/505fa4ea976e48f19f058f1285520910
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Phosphorylation of the mitochondrial autophagy receptor Nix enhances its interaction with LC3 proteins

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