Butler, Marion P. and Hanly, Jennifer A. and Moynagh, Paul N. (2007) Kinase-active Interleukin-1 Receptor-associated Kinases Promote Polyubiquitination and Degradation of the Pellino Family : direct evidence for PELLINO proteins being ubiquitin-protein isopeptide ligases. Journal of Biological Chemistry, 282 (41). pp. 29729-29737. ISSN 0021-9258

Preview

Download (448kB) | Preview

Abstract

Members of the Pellino family are interleukin-1 receptor-associated kinase (IRAK)-interacting proteins that possess RINGlike domains. The presence of these domains led to the suggestion that Pellino proteins are ubiquitin-protein isopeptide ligases (E3). However, no conclusive data currently exist to prove this proposal. This study provides the first direct evidence that Pellino proteins possess E3 activity. Recombinant forms of Pellino1 and Pellino2 and both spliced variants of Pellino3 are shown in an in vitro ubiquitination assay to be E3 ligases that catalyze Lys63-linked polyubiquitination, with Pellino3 exhibiting the greatest ligase activity. Whereas the Pellino proteins cause polyubiquitination of IRAK-1, we also show that kinaseactive members of the IRAK family (IRAK-1 and IRAK-4) promote reciprocal polyubiquitination of the Pellino proteins and that this is associated with IRAK-induced degradation of the Pellino family. In contrast, IRAK-2 (which lacks a functional kinase domain) and kinase-dead forms of IRAK-1 and IRAK-4 fail to degrade the Pellino proteins. We show that these kinaseinactive IRAK proteins can associate with Pellino proteins, thus excluding the possibility that their inability to regulate Pellino degradation is due to lack of association with the Pellino proteins. The physiological relevance of IRAK-induced degradation of Pellino proteins is confirmed by the demonstration that lipopolysaccharide causes degradation of endogenous forms of Pellino3 in peripheral blood mononuclear cells. In summary, this study not only demonstrates Pellino proteins to be E3 ligases that can catalyze Lys63-linked polyubiquitination but also shows bidirectional signaling between the IRAK and Pellino families and highlights a novel function for IRAK kinase activity.

Item Type:	Article
Additional Information:	This research was originally published in the Journal of Biological Chemistry. Butler, M.P., J.A. Hanly and P.N. Moynagh (2007) 'Kinase-active Interleukin-1 Receptor-associated Kinases Promote Polyubiquitination and Degradation of the Pellino Family: Direct evidence for pellino proteins being ubiquitin-protein isopeptide ligases'. Journal of Biological Chemistry, 282 :29729-29737
Keywords:	Kinase-active; interleukin-1; receptor-associated kinases; polyubiquitination; Pellino family; PELLINO proteins; ubiquitin-protein isopeptide ligases;
Academic Unit:	Faculty of Science and Engineering > Biology
Item ID:	7191
Identification Number:	https://doi.org/10.1074/jbc.M704558200
Depositing User:	Professor Paul Moynagh
Date Deposited:	15 Jul 2016 15:51
Journal or Publication Title:	Journal of Biological Chemistry
Publisher:	American Society for Biochemistry and Molecular Biology
Refereed:	Yes
Funders:	Health Research Board (HRB), Enterprise Ireland (EI)
URI:	Publisher
Use Licence:	This item is available under a Creative Commons Attribution Non Commercial Share Alike Licence (CC BY-NC-SA). Details of this licence are available here

Repository Staff Only(login required)

Item control page

Downloads

Origin of downloads