Published in

Wiley Open Access, Molecular Systems Biology, 1(5), p. 295, 2009

DOI: 10.1038/msb.2009.55

Wiley Open Access, Molecular Systems Biology, 1(5), p. 317, 2009

DOI: 10.1038/msb.2009.76

Links

Tools

Export citation

Search in Google Scholar

A comprehensive framework of E2–RING E3 interactions of the human ubiquitin–proteasome system

Journal article published in 2009 by Sjoerd J. L. van Wijk, Sjoerd J. de Vries, Patrick Kemmeren, Anding Huang, Rolf Boelens, Alexandre M. J. J. Bonvin ORCID, H. Th Marc Timmers
This paper is made freely available by the publisher.
This paper is made freely available by the publisher.

Full text: Download

Green circle
Preprint: archiving allowed
Upload
Green circle
Postprint: archiving allowed
Upload
Green circle
Published version: archiving allowed
Upload
Policy details
Data provided by SHERPA/RoMEO

Abstract

Covalent attachment of ubiquitin to substrates is crucial to protein degradation, transcription regulation and cell signalling. Highly specific interactions between ubiquitin-conjugating enzymes (E2) and ubiquitin protein E3 ligases fulfil essential roles in this process. We performed a global yeast-two hybrid screen to study the specificity of interactions between catalytic domains of the 35 human E2s with 250 RING-type E3s. Our analysis showed over 300 high-quality interactions, uncovering a large fraction of new E2–E3 pairs. Both within the E2 and the E3 cohorts, several members were identified that are more versatile in their interaction behaviour than others. We also found that the physical interactions of our screen compare well with reported functional E2–E3 pairs in in vitro ubiquitination experiments. For validation we confirmed the interaction of several versatile E2s with E3s in in vitro protein interaction assays and we used mutagenesis to alter the E3 interactions of the E2 specific for K63 linkages, UBE2N(Ubc13), towards the K48-specific UBE2D2(UbcH5B). Our data provide a detailed, genome-wide overview of binary E2–E3 interactions of the human ubiquitination system.