Structural basis for feedforward control in the PINK1/Parkin pathway. (2nd May 2022)

• Record Type: Journal Article
• Authors: Sauvé, Véronique; Sung, George; MacDougall, Emma J; Kozlov, Guennadi; Saran, Anshu; Fakih, Rayan; Fon, Edward A; Gehring, Kalle
• Is Part Of: EMBO journal. Volume 41:Number 12(2022)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2022-05-02
• Subjects: autophagy -- mitophagy -- open‐loop control -- Parkinson's disease -- ubiquitin; Molecular biology -- Periodicals; 572.805
• DOI: 10.15252/embj.2021109460
• Languages: English
• ISSNs: 0261-4189

Abstract

Abstract: PINK1 and parkin constitute a mitochondrial quality control system mutated in Parkinson's disease. PINK1, a kinase, phosphorylates ubiquitin to recruit parkin, an E3 ubiquitin ligase, to mitochondria. PINK1 controls both parkin localization and activity through phosphorylation of both ubiquitin and the ubiquitin‐like (Ubl) domain of parkin. Here, we observed that phospho‐ubiquitin can bind to two distinct sites on parkin, a high‐affinity site on RING1 that controls parkin localization and a low‐affinity site on RING0 that releases parkin autoinhibition. Surprisingly, ubiquitin vinyl sulfone assays, ITC, and NMR titrations showed that the RING0 site has higher affinity for phospho‐ubiquitin than phosphorylated Ubl in trans . We observed parkin activation by micromolar concentrations of tetra‐phospho‐ubiquitin chains that mimic mitochondria bearing multiple phosphorylated ubiquitins. A chimeric form of parkin with the Ubl domain replaced by ubiquitin was readily activated by PINK1 phosphorylation. In all cases, mutation of the binding site on RING0 abolished parkin activation. The feedforward mechanism of parkin activation confers robustness and rapidity to the PINK1‐parkin pathway and likely represents an intermediate step in its evolutionary development. Synopsis: The ubiquitin ligase parkin protects against Parkinson's disease by binding and ubiquitinating damaged mitochondria. Here, cellular, biochemical and biophysical results identify a novel feedforward mechanism involving direct parkin activation without phosphorylation by the protein kinase PINK1. Parkin has two binding sites for phosphorylated ubiquitin (pUb) The first binding site recruits parkin to damaged mitochondria The second site binds either pUb or the phosphorylated parkin Ubl domain, to switch on ubiquitin ligase activity Abstract : Two distinct phospho‐ubiquitin binding sites differentially control mitochondrial recruitment and activation of the parkin E3 ligase.