2. Academic Validation
  3. Ubiquitin is directly linked via an ester to protein-conjugated mono-ADP-ribose

Ubiquitin is directly linked via an ester to protein-conjugated mono-ADP-ribose

  • EMBO J. 2025 Feb 25. doi: 10.1038/s44318-025-00391-7.
Daniel S Bejan 1,  Rachel E Lacoursiere 2,  Jonathan N Pruneda 3 4 5,  Michael S Cohen 6 7
Affiliations

Affiliations

  • 1 Department of Chemical Physiology and Biochemistry, Oregon Health & Science University, Portland, OR, 97239, USA.
  • 2 Department of Molecular Microbiology and Immunology, Oregon Health & Science University, Portland, OR, 97239, USA.
  • 3 Department of Chemical Physiology and Biochemistry, Oregon Health & Science University, Portland, OR, 97239, USA. pruneda@ohsu.edu.
  • 4 Department of Molecular Microbiology and Immunology, Oregon Health & Science University, Portland, OR, 97239, USA. pruneda@ohsu.edu.
  • 5 Knight Cancer Institute, Oregon Health & Science University, Portland, OR, 97239, USA. pruneda@ohsu.edu.
  • 6 Department of Chemical Physiology and Biochemistry, Oregon Health & Science University, Portland, OR, 97239, USA. cohenmic@ohsu.edu.
  • 7 Knight Cancer Institute, Oregon Health & Science University, Portland, OR, 97239, USA. cohenmic@ohsu.edu.
PMID: 40000907 DOI: 10.1038/s44318-025-00391-7
Abstract

The prevailing view on post-translational modifications (PTMs) is that a single amino acid is modified with a single PTM at any given time. However, recent work has demonstrated crosstalk between different PTMs, some occurring on the same residue. Such interplay is seen with ADP-ribosylation and ubiquitylation. For example, DELTEX E3 Ligases were reported to ubiquitylate a hydroxyl group on free NAD+ and ADP-ribose in vitro, generating a noncanonical ubiquitin ester-linked species. In this report, we show, for the first time, that this dual PTM occurs in cells on mono-ADP-ribosylated (MARylated) PARP10 on Glu/Asp sites to form a MAR ubiquitin ester. We call this process mono-ADP-ribosyl ubiquitylation or MARUbylation. Using chemical and enzymatic treatments, including a newly characterized bacterial Deubiquitinase with esterase-specific activity, we discovered that multiple PARPs are MARUbylated and extended with K11-linked polyubiquitin chains when exogenously expressed. Finally, we show that in response to type I interferon stimulation, MARUbylation can occur endogenously on PARP targets. Thus, MARUbylation represents a new dual PTM that broadens our understanding of the function of PARP-mediated ADP-ribosylation in cells.

Keywords

ADP-ribosylation; Interferon Response; PARPs; Post-translational Modifications; Ubiquitylation.

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