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Identification of small ubiquitin-like modifier substrates with diverse functions using the Xenopus egg extract system.
Ma L
,
Aslanian A
,
Sun H
,
Jin M
,
Shi Y
,
Yates JR
,
Hunter T
.
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Post-translational modification by SUMO is a highly conserved pathway in eukaryotes that plays very important regulatory roles in many cellular processes. Deregulation of the SUMO pathway contributes to the development and progression of many diseases including cancer. Therefore, identifying additional SUMO substrates and studying how their cellular and biological functions are regulated by sumoylation should provide new insights. Our studies showed that sumoylation activity was significant in Xenopus egg extracts, and that a high level of sumoylation was associated with sperm chromatin when SUMO was incubated with Xenopus egg extracts. By isolating SUMO-conjugated substrates using His-tagged SUMO1 or SUMO2 proteins under denaturing conditions, we identified 346 proteins by mass spectrometry analysis that were not present in control pull-downs. Among them, 167 proteins were identified from interphase egg extracts, 86 proteins from mitotic phase egg extracts, and 93 proteins from both. Thirty-three proteins were pulled down by SUMO1, 85 proteins by SUMO2, and 228 proteins by both. We validated the sumoylation of five candidates, CKB, ATXN10, BTF3, HABP4, and BZW1, by co-transfecting them along with SUMO in HEK293T cells. Gene ontology analysis showed that SUMO substrates identified in this study were involved in diverse biological processes. Additionally, SUMO substrates identified from different cell cycle stages or pulled down by different SUMO homologs were enriched for distinct cellular components and functional categories. Our results comprehensively profile the sumoylation occurring in the Xenopus egg extract system.
Alarcon-Vargas,
SUMO in cancer--wrestlers wanted.
2002, Pubmed
Alarcon-Vargas,
SUMO in cancer--wrestlers wanted.
2002,
Pubmed
Ayaydin,
Distinct in vivo dynamics of vertebrate SUMO paralogues.
2004,
Pubmed
Azuma,
SUMO-2/3 regulates topoisomerase II in mitosis.
2003,
Pubmed
,
Xenbase
Bessman,
The creatine-creatine phosphate energy shuttle.
1985,
Pubmed
Bonne-Andrea,
SUMO2/3 modification of cyclin E contributes to the control of replication origin firing.
2013,
Pubmed
,
Xenbase
Bruderer,
Purification and identification of endogenous polySUMO conjugates.
2011,
Pubmed
Creton,
SnapShot: The SUMO system.
2010,
Pubmed
Dawlaty,
Resolution of sister centromeres requires RanBP2-mediated SUMOylation of topoisomerase IIalpha.
2008,
Pubmed
Eng,
An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein database.
1994,
Pubmed
Flotho,
Sumoylation: a regulatory protein modification in health and disease.
2013,
Pubmed
Franceschini,
STRING v9.1: protein-protein interaction networks, with increased coverage and integration.
2013,
Pubmed
Galisson,
A novel proteomics approach to identify SUMOylated proteins and their modification sites in human cells.
2011,
Pubmed
Geiss-Friedlander,
Concepts in sumoylation: a decade on.
2007,
Pubmed
Guo,
Proteomic analysis of SUMO4 substrates in HEK293 cells under serum starvation-induced stress.
2005,
Pubmed
Hannak,
Investigating mitotic spindle assembly and function in vitro using Xenopus laevis egg extracts.
2006,
Pubmed
,
Xenbase
Hayashi,
Ubc9 is essential for viability of higher eukaryotic cells.
2002,
Pubmed
Huang,
Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources.
2009,
Pubmed
Ishihama,
Exponentially modified protein abundance index (emPAI) for estimation of absolute protein amount in proteomics by the number of sequenced peptides per protein.
2005,
Pubmed
Johnson,
The ubiquitin-like protein Smt3p is activated for conjugation to other proteins by an Aos1p/Uba2p heterodimer.
1997,
Pubmed
Joseph,
SUMO-1 targets RanGAP1 to kinetochores and mitotic spindles.
2002,
Pubmed
Kessler,
A SUMOylation-dependent transcriptional subprogram is required for Myc-driven tumorigenesis.
2012,
Pubmed
Kim,
Systematic and quantitative assessment of the ubiquitin-modified proteome.
2011,
Pubmed
Knuesel,
A method of mapping protein sumoylation sites by mass spectrometry using a modified small ubiquitin-like modifier 1 (SUMO-1) and a computational program.
2005,
Pubmed
Leach,
Ubiquitin/SUMO modification of PCNA promotes replication fork progression in Xenopus laevis egg extracts.
2005,
Pubmed
,
Xenbase
Lohka,
Induction of nuclear envelope breakdown, chromosome condensation, and spindle formation in cell-free extracts.
1985,
Pubmed
,
Xenbase
Ma,
Requirement for Nudel and dynein for assembly of the lamin B spindle matrix.
2009,
Pubmed
,
Xenbase
Mahajan,
Molecular characterization of the SUMO-1 modification of RanGAP1 and its role in nuclear envelope association.
1998,
Pubmed
Matic,
Site-specific identification of SUMO-2 targets in cells reveals an inverted SUMOylation motif and a hydrophobic cluster SUMOylation motif.
2010,
Pubmed
McDonald,
MS1, MS2, and SQT-three unified, compact, and easily parsed file formats for the storage of shotgun proteomic spectra and identifications.
2004,
Pubmed
Medema,
Boosting and suppressing mitotic phosphorylation.
2011,
Pubmed
Melchior,
SUMO-1 and p53.
2002,
Pubmed
Mi,
PANTHER in 2013: modeling the evolution of gene function, and other gene attributes, in the context of phylogenetic trees.
2013,
Pubmed
Mudunuri,
bioDBnet: the biological database network.
2009,
Pubmed
März,
Ataxin-10, the spinocerebellar ataxia type 10 neurodegenerative disorder protein, is essential for survival of cerebellar neurons.
2004,
Pubmed
Panse,
A proteome-wide approach identifies sumoylated substrate proteins in yeast.
2004,
Pubmed
Peng,
Evaluation of multidimensional chromatography coupled with tandem mass spectrometry (LC/LC-MS/MS) for large-scale protein analysis: the yeast proteome.
2003,
Pubmed
Powers,
Preparation and use of interphase Xenopus egg extracts.
2001,
Pubmed
,
Xenbase
Psakhye,
Protein group modification and synergy in the SUMO pathway as exemplified in DNA repair.
2012,
Pubmed
Ryu,
PIASy mediates SUMO-2/3 conjugation of poly(ADP-ribose) polymerase 1 (PARP1) on mitotic chromosomes.
2010,
Pubmed
,
Xenbase
Saitoh,
Ubc9p and the conjugation of SUMO-1 to RanGAP1 and RanBP2.
1998,
Pubmed
,
Xenbase
Saitoh,
Functional heterogeneity of small ubiquitin-related protein modifiers SUMO-1 versus SUMO-2/3.
2000,
Pubmed
Schimmel,
Uncovering SUMOylation dynamics during cell-cycle progression reveals FoxM1 as a key mitotic SUMO target protein.
2014,
Pubmed
Shiio,
Histone sumoylation is associated with transcriptional repression.
2003,
Pubmed
Sumara,
E3 ubiquitin ligases and mitosis: embracing the complexity.
2008,
Pubmed
Sun,
Conserved function of RNF4 family proteins in eukaryotes: targeting a ubiquitin ligase to SUMOylated proteins.
2007,
Pubmed
Sun,
Poly-small ubiquitin-like modifier (PolySUMO)-binding proteins identified through a string search.
2012,
Pubmed
Tabb,
DTASelect and Contrast: tools for assembling and comparing protein identifications from shotgun proteomics.
2002,
Pubmed
Tsai,
A mitotic lamin B matrix induced by RanGTP required for spindle assembly.
2006,
Pubmed
,
Xenbase
Ubersax,
Targets of the cyclin-dependent kinase Cdk1.
2003,
Pubmed
Vertegaal,
Distinct and overlapping sets of SUMO-1 and SUMO-2 target proteins revealed by quantitative proteomics.
2006,
Pubmed
Vethantham,
In vitro sumoylation of recombinant proteins and subsequent purification for use in enzymatic assays.
2009,
Pubmed
Vigodner,
Localization and identification of sumoylated proteins in human sperm: excessive sumoylation is a marker of defective spermatozoa.
2013,
Pubmed
Watts,
The role of SUMO in chromosome segregation.
2007,
Pubmed
Xu,
Global analysis of lysine ubiquitination by ubiquitin remnant immunoaffinity profiling.
2010,
Pubmed
Zhang,
SUMO-2/3 modification and binding regulate the association of CENP-E with kinetochores and progression through mitosis.
2008,
Pubmed
,
Xenbase
Zhao,
Sumoylation regulates diverse biological processes.
2007,
Pubmed
Zheng,
Sequencing and expression of complementary DNA for the general transcription factor BTF3.
1990,
Pubmed
Zhou,
Global analyses of sumoylated proteins in Saccharomyces cerevisiae. Induction of protein sumoylation by cellular stresses.
2004,
Pubmed
Zybailov,
Statistical analysis of membrane proteome expression changes in Saccharomyces cerevisiae.
2006,
Pubmed
