Identification of RNF114 as ADPr-Ub reader through non-hydrolysable ubiquitinated ADP-ribose

  • Max S. Kloet
  • , Chatrin Chatrin*
  • , Rishov Mukhopadhyay
  • , Bianca D.M. van Tol
  • , Rebecca Smith
  • , Sarah A. Rotman
  • , Rayman T.N. Tjokrodirijo
  • , Kang Zhu
  • , Andrii Gorelik
  • , Lucy Maginn
  • , Paul R. Elliott
  • , Peter A. van Veelen
  • , Dragana Ahel
  • , Ivan Ahel*
  • , Gerbrand J. van der Heden van Noort*
  • *Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

Crosstalk between the post-translational modification processes of ubiquitination and ADP-ribosylation occurs in DNA-damage- and immune-responses, in addition the physical linkage of ADP-ribose and ubiquitin is found during bacterial infection. Here, we study the ubiquitination of ADP-ribose mediated by human Deltex E3 ligases and the subsequent fate of the formed hybrid post-translational modification. We prepare a non-hydrolysable ADPr-Ub probe that we employ in a proteomics approach and identify RNF114 as an interacting protein. Using biophysical and biochemical experiments, we validate that RNF114 preferentially interacts with ubiquitinated ADP-ribose over non-modified ubiquitin. Subsequently, RNF114 can elongate the ubiquitinated ADP-ribose with a K11-linked ubiquitin chain. Using domain deletion analysis, we pinpoint the tandem zinc fingers and ubiquitin interacting motif (ZnF2 + ZnF3+UIM) domains of RNF114 to be crucial for recognising ubiquitinated ADP-ribose. Moreover, these domains are essential for the recruitment of RNF114 to the sites of laser-induced DNA damage.

Original languageEnglish
Article number6319
JournalNature Communications
Volume16
Issue number1
DOIs
StatePublished - Dec 2025
Externally publishedYes

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