PINK1 phosphorylates ubiquitin to activate parkin E3 ubiquitin ligase activity

Lesley A. Kane, Michael Lazarou, Adam I. Fogel, Yan Li, Koji Yamano, Shireen A. Sarraf, Soojay Banerjee, Richard J. Youle

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553 Citations (Scopus)

Abstract

PINK1 kinase activates the E3 ubiquitin ligase Parkin to induce selective autophagy of damaged mitochondria. However, it has been unclear how PINK1 activates and recruits Parkin to mitochondria. Although PINK1 phosphorylates Parkin, other PINK1 substrates appear to activate Parkin, as the mutation of all serine and threonine residues conserved between Drosophila and human, including Parkin S65, did not wholly impair Parkin translocation to mitochondria. Using mass spectrometry, we discovered that endogenous PINK1 phosphorylated ubiquitin at serine 65, homologous to the site phosphorylated by PINK1 in Parkin's ubiquitin-like domain. Recombinant TcPINK1 directly phosphorylated ubiquitin and phosphoubiquitin activated Parkin E3 ubiquitin ligase activity in cell-free assays. In cells, the phosphomimetic ubiquitin mutant S65D bound and activated Parkin. Furthermore, expression of ubiquitin S65A, a mutant that cannot be phosphorylated by PINK1, inhibited Parkin translocation to damaged mitochondria. These results explain a feedforward mechanism of PINK1-mediated initiation of Parkin E3 ligase activity.

Original languageEnglish
Pages (from-to)143-153
Number of pages11
JournalJournal of Cell Biology
Volume205
Issue number2
DOIs
Publication statusPublished - 28 Apr 2014
Externally publishedYes

Cite this

Kane, L. A., Lazarou, M., Fogel, A. I., Li, Y., Yamano, K., Sarraf, S. A., Banerjee, S., & Youle, R. J. (2014). PINK1 phosphorylates ubiquitin to activate parkin E3 ubiquitin ligase activity. Journal of Cell Biology, 205(2), 143-153. https://doi.org/10.1083/jcb.201402104