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Uncovering the SUMOylation and ubiquitylation crosstalk in human cells using sequential peptide immunopurification.

Authors :
Lamoliatte F
McManus FP
Maarifi G
Chelbi-Alix MK
Thibault P
Source :
Nature communications [Nat Commun] 2017 Jan 18; Vol. 8, pp. 14109. Date of Electronic Publication: 2017 Jan 18.
Publication Year :
2017

Abstract

Crosstalk between the SUMO and ubiquitin pathways has recently been reported. However, no approach currently exists to determine the interrelationship between these modifications. Here, we report an optimized immunoaffinity method that permits the study of both protein ubiquitylation and SUMOylation from a single sample. This method enables the unprecedented identification of 10,388 SUMO sites in HEK293 cells. The sequential use of SUMO and ubiquitin remnant immunoaffinity purification facilitates the dynamic profiling of SUMOylated and ubiquitylated proteins in HEK293 cells treated with the proteasome inhibitor MG132. Quantitative proteomic analyses reveals crosstalk between substrates that control protein degradation, and highlights co-regulation of SUMOylation and ubiquitylation levels on deubiquitinase enzymes and the SUMOylation of proteasome subunits. The SUMOylation of the proteasome affects its recruitment to promyelocytic leukemia protein (PML) nuclear bodies, and PML lacking the SUMO interacting motif fails to colocalize with SUMOylated proteasome further demonstrating that this motif is required for PML catabolism.

Details

Language :
English
ISSN :
2041-1723
Volume :
8
Database :
MEDLINE
Journal :
Nature communications
Publication Type :
Academic Journal
Accession number :
28098164
Full Text :
https://doi.org/10.1038/ncomms14109