Ubiquitin Ser65 phosphorylation affects ubiquitin structure, chain assembly and hydrolysis

Abstract

The protein kinase PINK1 was recently shown to phosphorylate ubiquitin (Ub) on Ser65, and phosphoUb activates the E3 ligase Parkin allosterically. Here, we show that PINK1 can phosphorylate every Ub in Ub chains. Moreover, Ser65 phosphorylation alters Ub structure, generating two conformations in solution. A crystal structure of the major conformation resembles Ub but has altered surface properties. NMR reveals a second phosphoUb conformation in which β5‐strand slippage retracts the C‐terminal tail by two residues into the Ub core. We further show that phosphoUb has no effect on E1‐mediated E2 charging but can affect discharging of E2 enzymes to form polyUb chains. Notably, UBE2R1‐ (CDC34), UBE2N/UBE2V1‐ (UBC13/UEV1A), TRAF6‐ and HOIP‐mediated chain assembly is inhibited by phosphoUb. While Lys63‐linked poly‐phosphoUb is recognized by the TAB2 NZF Ub binding domain (UBD), 10 out of 12 deubiquitinases (DUBs), including USP8, USP15 and USP30, are impaired in hydrolyzing phosphoUb chains. Hence, Ub phosphorylation has repercussions for ubiquitination and deubiquitination cascades beyond Parkin activation and may provide an independent layer of regulation in the Ub system.

Cited by papers we hold 18

• Elevated ubiquitin phosphorylation by PINK1 contributes to proteasomal impairment and promotes neurodegeneration · 2025
  Not checked
• The mitophagy pathway and its implications in human diseases · 2023
  No notice
• Mitophagy: Molecular Mechanisms, New Concepts on Parkin Activation and the Emerging Role of AMPK/ULK1 Axis · 2021
  No notice
• Molecular mechanisms and physiological functions of mitophagy · 2021
  No notice
• Activation mechanism of PINK1 · 2021
  Corrected
• Selective autophagy of intracellular organelles: Recent research advances · 2021
  No notice
• MitophAging: Mitophagy in Aging and Disease · 2020
  No notice
• The role of ubiquitination in tumorigenesis and targeted drug discovery · 2020
  No notice
• Mitophagy: An Emerging Role in Aging and Age-Associated Diseases · 2020
  No notice
• USP30 sets a trigger threshold for PINK1–PARKIN amplification of mitochondrial ubiquitylation · 2020
  No notice
• Dual role of USP30 in controlling basal pexophagy and mitophagy · 2018
  No notice
• PTEN-L is a novel protein phosphatase for ubiquitin dephosphorylation to inhibit PINK1–Parkin-mediated mitophagy · 2018
  Corrected
• Mfn2 ubiquitination by PINK1/parkin gates the p97-dependent release of ER from mitochondria to drive mitophagy · 2018
  No notice
• PINK1 import regulation; a fine system to convey mitochondrial stress to the cytosol · 2018
  No notice
• Ubiquitin Linkage-Specific Affimers Reveal Insights into K6-Linked Ubiquitin Signaling · 2017
  No notice
• The recognition of ubiquitinated proteins by the proteasome · 2016
  No notice
• Ubiquitin modifications · 2016
  No notice
• Phosphoproteomic screening identifies Rab GTPases as novel downstream targets of PINK1 · 2015
  No notice

Reads

A person counts once a day per paper. We keep no cookie and no address: a reader is a hash that changes every day. Any month under five reads shows no number.