Mapping of SUMO sites and analysis of SUMOylation changes induced by external stimuli

Francis Impens; Lilliana Radoshevich; Pascale Cossart; David Ribet

doi:10.1073/pnas.1413825111

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Mapping of SUMO sites and analysis of SUMOylation changes induced by external stimuli

Journal article

Open access

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Mapping of SUMO sites and analysis of SUMOylation changes induced by external stimuli

Francis Impens, Lilliana Radoshevich, Pascale Cossart and David Ribet

Proceedings of the National Academy of Sciences - PNAS, Vol.111(34), pp.12432-12437

08/26/2014

DOI: 10.1073/pnas.1413825111

PMCID: PMC4151716

PMID: 25114211

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Abstract

SUMOylation is an essential ubiquitin-like modification involved in important biological processes in eukaryotic cells. Identification of small ubiquitin-related modifier (SUMO)-conjugated residues in proteins is critical for understanding the role of SUMOylation but remains experimentally challenging. We have set up a powerful and high-throughput method combining quantitative proteomics and peptide immunocapture to map SUMOylation sites and have analyzed changes in SUMOylation in response to stimuli. With this technique we identified 295 SUMO1 and 167 SUMO2 sites on endogenous substrates of human cells. We further used this strategy to characterize changes in SUMOylation induced by listeriolysin O, a bacterial toxin that impairs the host cell SUMOylation machinery, and identified several classes of host proteins specifically deSUMOylated in response to this toxin. Our approach constitutes an unprecedented tool, broadly applicable to various SUMO-regulated cellular processes in health and disease.

Recombinant Proteins - metabolism

Amino Acid Sequence

Small Ubiquitin-Related Modifier Proteins - metabolism

Humans

Bacterial Toxins - toxicity

Molecular Sequence Data

Recombinant Proteins - chemistry

Peptide Mapping - methods

Recombinant Proteins - genetics

Sumoylation - drug effects

Small Ubiquitin-Related Modifier Proteins - genetics

SUMO-1 Protein - chemistry

SUMO-1 Protein - genetics

Small Ubiquitin-Related Modifier Proteins - chemistry

Hemolysin Proteins - toxicity

SUMO-1 Protein - metabolism

HeLa Cells

Proteomics - methods

Binding Sites

Heat-Shock Proteins - toxicity

Details

Title: Subtitle: Mapping of SUMO sites and analysis of SUMOylation changes induced by external stimuli
Creators: Francis Impens - Unité des Interactions Bactéries-Cellules, Institut Pasteur, F-75015 Paris, France; Institut National de la Santé et de la Recherche Médicale, Unité 604, F-75015 Paris, France; and Institut National de la Recherche Agronomique, Unité sous-contrat 2020, F-75015 Paris, France
Lilliana Radoshevich - Unité des Interactions Bactéries-Cellules, Institut Pasteur, F-75015 Paris, France; Institut National de la Santé et de la Recherche Médicale, Unité 604, F-75015 Paris, France; and Institut National de la Recherche Agronomique, Unité sous-contrat 2020, F-75015 Paris, France
Pascale Cossart - Unité des Interactions Bactéries-Cellules, Institut Pasteur, F-75015 Paris, France; Institut National de la Santé et de la Recherche Médicale, Unité 604, F-75015 Paris, France; and Institut National de la Recherche Agronomique, Unité sous-contrat 2020, F-75015 Paris, France pcossart@pasteur.fr david.ribet@pasteur.fr
David Ribet - Unité des Interactions Bactéries-Cellules, Institut Pasteur, F-75015 Paris, France; Institut National de la Santé et de la Recherche Médicale, Unité 604, F-75015 Paris, France; and Institut National de la Recherche Agronomique, Unité sous-contrat 2020, F-75015 Paris, France pcossart@pasteur.fr david.ribet@pasteur.fr
Resource Type: Journal article
Publication Details: Proceedings of the National Academy of Sciences - PNAS, Vol.111(34), pp.12432-12437
DOI: 10.1073/pnas.1413825111
PMID: 25114211
PMCID: PMC4151716
NLM abbreviation: Proc Natl Acad Sci U S A
ISSN: 0027-8424
eISSN: 1091-6490
Publisher: National Academy of Sciences; United States
Language: English
Date published: 08/26/2014
Academic Unit: Molecular Physiology and Biophysics; Microbiology and Immunology
Record Identifier: 9984025348202771

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