Formation of functional E3 ligase complexes with UBC2 and UEV1 of Leishmania mexicana

Rebecca J. Burge, Katie H. Jameson, Vincent Geoghegan, Adam A. Dowle, Jeremy C. Mottram*, Anthony J. Wilkinson

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


In eukaryotic cells, molecular fate and cellular responses are shaped by multicomponent enzyme systems which reversibly attach ubiquitin and ubiquitin-like modifiers to target proteins. The extent of the ubiquitin proteasome system in Leishmania mexicana and its importance for parasite survival has recently been established through deletion mutagenesis and life-cycle phenotyping studies. The ubiquitin conjugating E2 enzyme UBC2, and the E2 enzyme variant UEV1, with which it forms a stable complex in vitro, were shown to be essential for the differentiation of promastigote parasites to the infectious amastigote form. To investigate further, we used immunoprecipitation of Myc-UBC2 or Myc-UEV1 to identify interacting proteins in L. mexicana promastigotes. The interactome of UBC2 comprises multiple ubiquitin-proteasome components including UEV1 and four RING E3 ligases, as well as potential substrates predicted to have roles in carbohydrate metabolism and intracellular trafficking. The smaller UEV1 interactome comprises six proteins, including UBC2 and shared components of the UBC2 interactome consistent with the presence of intracellular UBC2-UEV1 complexes. Recombinant RING1, RING2 and RING4 E3 ligases were shown to support ubiquitin transfer reactions involving the E1, UBA1a, and UBC2 to available substrate proteins or to unanchored ubiquitin chains. These studies define additional components of a UBC2-dependent ubiquitination pathway shown previously to be essential for promastigote to amastigote differentiation.

Original languageEnglish
Article number111619
Number of pages11
Early online date29 Mar 2024
Publication statusPublished - Jun 2024

Bibliographical note

© 2024 The Authors. Published by Elsevier B.V.


  • Interactome
  • Leishmania
  • Proteomics
  • Ubiquitin
  • Ubiquitin ligase
  • Ubiquitin-conjugating enzyme (E2 enzyme)

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