title: Systematic identification and analysis of mammalian small ubiquitin-like modifier substrates.
abstract: Small ubiquitin-like modifier (SUMO) regulates diverse cellular processes through its reversible, covalent attachment to target proteins. Many SUMO substrates are involved in transcription and chromatin structure. Sumoylation appears to regulate the functions of target proteins by changing their subcellular localization, increasing their stability, and/or mediating their binding to other proteins. Using an in vitro expression cloning approach, we have identified 40 human SUMO1 substrates. The spectrum of human SUMO1 substrates identified in our screen suggests general roles of sumoylation in transcription, chromosome structure, and RNA processing. We have validated the sumoylation of 24 substrates in living cells. Analysis of this panel of SUMO substrates leads to the following observations. 1) Sumoylation is more efficient in vitro than in living cells. Polysumoylation occurs on several substrates in vitro. 2) SUMO isopeptidases have little substrate specificity. 3) The SUMO ligases, PIAS1 and PIASxbeta, have broader substrate specificities than does PIASy. 4) Although SUMO1 and SUMO2 are equally efficiently conjugated to a given substrate in vitro, SUMO1 conjugation is more efficient in vivo. 5) Most SUMO substrates localize to the nucleus, and sumoylation does not generally affect their subcellular localization. Therefore, sumoylation appears to regulate the functions of its substrates through multiple, context-dependent mechanisms.
authors.affiliation: Department of Pharmacology, the University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA.
authors.forename: Christian B
authors.initials: CB
authors.lastname: Gocke
authors.forename: Hongtao
authors.initials: H
authors.lastname: Yu
authors.forename: Jungseog
authors.initials: J
authors.lastname: Kang
mesh_terms: D004247:DNA / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry; D051741:Kruppel-Like Transcription Factors; D008239:Lysine / Q000737:chemistry; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000378:metabolism; D000075223:Poly-ADP-Ribose Binding Proteins; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D050828:Protein Inhibitors of Activated STAT; D011506:Proteins / Q000737:chemistry; D012313:RNA / Q000737:chemistry; D025842:SUMO-1 Protein; D025841:Small Ubiquitin-Related Modifier Proteins / Q000378:metabolism*; D013379:Substrate Specificity; D013997:Time Factors; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism; D014162:Transfection
chemical_list: D018076:DNA, Complementary; D004268:DNA-Binding Proteins; C095239:ETV1 protein, human; D047908:Intracellular Signaling Peptides and Proteins; D051741:Kruppel-Like Transcription Factors; C485891:PIAS4 protein, human; D000075223:Poly-ADP-Ribose Binding Proteins; D050828:Protein Inhibitors of Activated STAT; D011506:Proteins; D025842:SUMO-1 Protein; C495501:SUMO1 protein, human; C117909:SUMO2 protein, human; D025841:Small Ubiquitin-Related Modifier Proteins; D014157:Transcription Factors; C107961:ZBTB17 protein, human; D012313:RNA; D004247:DNA; D008239:Lysine