Comprehensive identification of SUMO2/3 targets and their dynamics during mitosis

Julie Schou; Christian D Kelstrup; Daniel G Hayward; Jesper V Olsen; Jakob Nilsson

doi:10.1371/journal.pone.0100692

Comprehensive identification of SUMO2/3 targets and their dynamics during mitosis

PLoS One. 2014 Jun 27;9(6):e100692. doi: 10.1371/journal.pone.0100692. eCollection 2014.

Authors

Julie Schou¹, Christian D Kelstrup¹, Daniel G Hayward¹, Jesper V Olsen¹, Jakob Nilsson¹

Affiliation

¹ The Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.

Abstract

During mitosis large alterations in cellular structures occur rapidly, which to a large extent is regulated by post-translational modification of proteins. Modification of proteins with the small ubiquitin-related protein SUMO2/3 regulates mitotic progression, but few mitotic targets have been identified so far. To deepen our understanding of SUMO2/3 during this window of the cell cycle, we undertook a comprehensive proteomic characterization of SUMO2/3 modified proteins in mitosis and upon mitotic exit. We developed an efficient tandem affinity purification strategy of SUMO2/3 modified proteins from mitotic cells. Combining this purification strategy with cell synchronization procedures and quantitative mass spectrometry allowed for the mapping of numerous novel targets and their dynamics as cells progressed out of mitosis. This identified RhoGDIα as a major SUMO2/3 modified protein, specifically during mitosis, mediated by the SUMO ligases PIAS2 and PIAS3. Our data provide a rich resource for further exploring the role of SUMO2/3 modifications in mitosis and cell cycle regulation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Amino Acid Substitution
Benzamides / pharmacology
Chromatography, High Pressure Liquid
HeLa Cells
Humans
Mitosis*
Molecular Sequence Data
Paclitaxel / pharmacology
Prometaphase
Proteomics
Quinazolines / pharmacology
RNA Interference
RNA, Small Interfering / metabolism
Small Ubiquitin-Related Modifier Proteins / chemistry
Small Ubiquitin-Related Modifier Proteins / genetics
Small Ubiquitin-Related Modifier Proteins / metabolism*
Sumoylation / drug effects
Tandem Mass Spectrometry
Ubiquitins / chemistry
Ubiquitins / genetics
Ubiquitins / metabolism*
rho Guanine Nucleotide Dissociation Inhibitor alpha / antagonists & inhibitors
rho Guanine Nucleotide Dissociation Inhibitor alpha / genetics
rho Guanine Nucleotide Dissociation Inhibitor alpha / metabolism
rho Guanine Nucleotide Dissociation Inhibitor beta / antagonists & inhibitors
rho Guanine Nucleotide Dissociation Inhibitor beta / genetics
rho Guanine Nucleotide Dissociation Inhibitor beta / metabolism

Substances

4-(4-(N-benzoylamino)anilino)-6-methoxy-7-(3-(1-morpholino)propoxy)quinazoline
Benzamides
Quinazolines
RNA, Small Interfering
SUMO2 protein, human
SUMO3 protein, human
Small Ubiquitin-Related Modifier Proteins
Ubiquitins
rho Guanine Nucleotide Dissociation Inhibitor alpha
rho Guanine Nucleotide Dissociation Inhibitor beta
Paclitaxel

Grants and funding

Lundbeck Foundation (www.lundbeckfonden.com) supported this work through a Junior Group Leader Fellowship to JN. The Novo Nordisk Foundation (www.novonordiskfonden.dk) supported this through institute funding for JN. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.