Assembly and structure of Lys33-linked polyubiquitin reveals distinct conformations

Yosua Adi Kristariyanto, Soo-Youn Choi, Syed Arif Abdul Rehman, Maria Stella Ritorto, David G. Campbell, Nicholas A. Morrice, Rachel Toth, Yogesh Kulathu (Lead / Corresponding author)

Research output: Contribution to journalArticlepeer-review

60 Citations (Scopus)
194 Downloads (Pure)


Ubiquitylation regulates a multitude of biological processes and this versatility stems from the ability of ubiquitin (Ub) to form topologically different polymers of eight different linkage types. Whereas some linkages have been studied in detail, other linkage types including Lys<sup>33</sup> -linked polyUb are poorly understood. In the present study, we identify an enzymatic system for the large-scale assembly of Lys<sup>33</sup> chains by combining the HECT (homologous to the E6-AP C-terminus) E3 ligase AREL1 (apoptosis-resistant E3 Ub protein ligase 1) with linkage selective deubiquitinases (DUBs). Moreover, this first characterization of the chain selectivity of AREL1 indicates its preference for assembling Lys<sup>33</sup> - and Lys<sup>11</sup> -linked Ub chains. Intriguingly, the crystal structure of Lys<sup>33</sup> -linked diUb reveals that it adopts a compact conformation very similar to that observed for Lys<sup>11</sup>- linked diUb. In contrast, crystallographic analysis of Lys<sup>33</sup> -linked triUb reveals a more extended conformation. These two distinct conformational states of Lys<sup>33</sup> -linked polyUb may be selectively recognized by Ub-binding domains (UBD) and enzymes of the Ub system. Importantly, our work provides a method to assemble Lys<sup>33</sup> -linked polyUb that will allow further characterization of this atypical chain type.

Original languageEnglish
Pages (from-to)345-352
Number of pages8
JournalBiochemical Journal
Issue number2
Publication statusPublished - 15 Apr 2015


  • Deubiquitinase
  • Homologous to the e6-apc-terminus (hect) e3 ligase
  • Polyubiquitin
  • Ubiquitin linkage

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology


Dive into the research topics of 'Assembly and structure of Lys33-linked polyubiquitin reveals distinct conformations'. Together they form a unique fingerprint.

Cite this