Unconventional initiation of PINK1/Parkin mitophagy by Optineurin

Nguyen, Thanh N.; Sawa-Makarska, Justyna; Khuu, Grace; Lam, Wai Kit; Adriaenssens, Elias; Fracchiolla, Dorotea; Shoebridge, Stephen; Bernklau, Daniel; Padman, Benjamin Scott; Skulsuppaisarn, Marvin; Lindblom, Runa; Martens, Sascha; Lazarou, Michael

doi:10.1016/j.molcel.2023.04.021

Cited by 34 publications

(25 citation statements)

References 101 publications

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“…2b and Extended Data Fig. 7h), and TBK1 has been shown to bind directly to PI3KC3-C1 28 . It will be interesting to determine if TBK1 uses its FIP200 arm-like domain to bypass the need for ULK1/2 in OPTN mitophagy.…”

Section: Discussionmentioning

confidence: 95%

“…Most, but not all, autophagy initiation pathways require ULK1/2. In OPTN-mediated Parkin-dependent mitophagy, TBK1 replaces the requirement for ULK1/2 28 . However, ATG13 and FIP200 are still essential for this pathway, as confirmed here by the phenotype observed for ATG13-binding deficient FIP200 mutations.…”

Section: Discussionmentioning

confidence: 99%

“…mCherry-tagged FIP200 wild type (WT) and mutants (V44D; V314D; V44D/V314D; L324D/R543/R548D; V44D/V314D/L324/R543/R548D were subcloned into pCDH1-CMV-MCS-SV40-hygromycin lentiviral vector using NheI and BamHI restriction enzyme sites. Generation of stable cell lines were previously described 31,52 and available at protocols.io (doi.org/10.17504/protocols.io.dm6gpjzm8gzp/v1). BFP-Parkin (Addgene, 186221) and pSu9-Halo-GFP (Addgene, 184905) were stably expressed in FIP200 KO cells using lentiviral transduction.…”

Section: Methodsmentioning

confidence: 99%

“…Mitophagy induction was carried out as described in refs. 31,54 . Briefly, 350,000 cells were seeded the day before the mitophagy induction in 6 well plates.…”

Section: Methodsmentioning

confidence: 99%

“…We therefore tested the function of both single and multisite FIP200 mutations. The first steps in PINK1/Parkin mitophagy can bypass ULK1 recruitment via the OPTN-TBK1-PI3KC3-C1 axis, while NDP52 can utilize either ULK1 or TBK1, 31 and therefore a full loss of function was not expected. Most FIP200 mutations did not lead to an evident loss of function.…”

Section: Mainmentioning

confidence: 99%

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Structure and activation of the human autophagy-initiating ULK1C:PI3KC3-C1 supercomplex

Chen

Ren

Cook

et al. 2023

Preprint

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The unc-51-like kinase protein kinase complex (ULK1C) and the class III phosphatidylinositol (PI) 3-kinase complex I (PI3KC3-C1) are the most upstream and central players in the initiation of macroautophagy. We found a direct physical interaction between the two complexes. The cryo-EM structures of the human ULK1C core and PI3KC3-C1 were determined at amino acid residue-level resolution, and used to interpret a moderate resolution structure of the ULK1C:PI3KC3-C1 supercomplex. The two complexes coassemble through extensive contacts between the FIP200 scaffold subunit of ULK1C and the VPS15 pseudokinase subunit of PI3KC3-C1. The presence of PI3KC3-C1 induces a rearrangement of ULK1C from a FIP200:ATG13:ULK1 2:1:1 to a 2:2:2 stoichiometry by dislocating an ATG13 loop (ATG13MR) from an inhibitory site on the dimeric FIP200 scaffold. This suggests a mechanism for the initiation of autophagy through PI3KC3-C1-induced dimerization of ULK1 as bound to FIP200, followed by an activating trans-autophosphorylation of ULK1.

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Section: Discussionmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…Mitophagy induction was carried out as described in refs. 31,54 . Briefly, 350,000 cells were seeded the day before the mitophagy induction in 6 well plates.…”

Section: Methodsmentioning

confidence: 99%

Section: Mainmentioning

confidence: 99%

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