Coupled monoubiquitylation of the co-E3 ligase DCNL1 by Ariadne RBR E3 ubiquitin ligases promotes cullin-RING ligase complex remodeling

Ian R. Kelsall, Yosua A. Kristariyanto, Axel Knebel, Nicola T. Wood, Yogesh Kulathu, Arno F. Alpi (Lead / Corresponding author)

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Cullin-RING E3 ubiquitin ligases (CRLs) are large and diverse multisubunit protein complexes that contribute to about one-fifth of ubiquitin-dependent protein turnover in cells. CRLs are activated by the attachment of the ubiquitin-like protein neural precursor cell expressed, developmentally down-regulated 8 (NEDD8) to the cullin subunits. This cullin neddylation is essential for a plethora of CRL-regulated cellular processes and vital for life. In mammals, neddylation is promoted by the five co-E3 ligases, defective in cullin neddylation 1 domain-containing 1-5 (DCNL1-5); however, their functional regulation within the CRL complex remains elusive. We found here that the ubiquitin associated (UBA)-containing DCNL1 is monoubiquitylated when bound to CRLs and that this monoubiquitylation depends on the CRL-associated Ariadne RBR ligases TRIAD1 (ARIH2) and HHARI (ARIH1), and strictly requires DCNL1's UBA domain. Reconstitution of DCNL1 monoubiquitylation in vitro revealed that autoubiquitylated TRIAD1 mediates binding to the UBA domain and subsequently promotes a single ubiquitin attachment to DCNL1 in a mechanism previously dubbed coupled monoubiquitylation. Moreover, we provide evidence that DCNL1 monoubiquitylation is required for efficient CRL activity, most likely by remodeling CRLs and their substrate receptors. Collectively, this work identifies DCNL1 as a critical target of Ariadne RBR ligases and coupled monoubiquitylation of DCNL1 as an integrated mechanism that affects CRL activity and client-substrate ubiquitylation at multiple levels.

Original languageEnglish
Pages (from-to)2651-2664
Number of pages14
JournalJournal of Biological Chemistry
Issue number8
Early online date26 Dec 2018
Publication statusPublished - 22 Feb 2019


  • ARIH1
  • ARIH2
  • NEDD8 E3 ligase
  • DCNL1
  • coupled monoubiquitylation
  • ubiquitylation (ubiquitination)
  • ubiquitin
  • cell signaling
  • post-translational modification (PTM)
  • ubiquitin ligase


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