title: Ubc9 fusion-directed SUMOylation identifies constitutive and inducible SUMOylation.
abstract: Constitutive and induced protein SUMOylation is involved in the regulation of a variety of cellular processes, such as regulation of gene expression and protein transport, and proceeds mainly in the nucleus of the cell. So far, several hundred SUMOylation targets have been identified, but presumably they represent only a part of the total of proteins which are regulated by SUMOylation. Here, we used the Ubc9 fusion-dependent SUMOylation system (UFDS) to screen for constitutive and induced SUMOylation of 46 randomly chosen proteins with proven or potential nuclear localization. Fourteen new UFDS-substrate proteins were identified of which eight could be demonstrated to be SUMOylated in a UFDS-independent manner in vivo. Of these, three were constitutively SUMOylated (FOS, CRSP9 and CDC37) while the remaining five substrates (CSNK2B, TAF10, HSF2BP, PSMC3 and DRG1) showed a stimulation-dependent SUMOylation induced by the MAP3 kinase MEKK1. Hence, UFDS is appropriate for the identification and characterization of constitutive and, more importantly, induced protein SUMOylation in vivo.
authors: Institut für Physiologische Chemie, Medizinische Hochschule Hannover, Carl-Neuberg Street, 1, 30625 Hannover, Germany.
authors: Astrid
authors: A
authors: Jakobs
authors: Fabian
authors: F
authors: Himstedt
authors: Martin
authors: M
authors: Funk
authors: Bernhard
authors: B
authors: Korn
authors: Matthias
authors: M
authors: Gaestel
authors: Rainer
authors: R
authors: Niedenthal
mesh_terms: D002460:Cell Line; D006801:Humans; D048728:MAP Kinase Kinase Kinase 1 / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D025841:Small Ubiquitin-Related Modifier Proteins / Q000378:metabolism*; D044763:Ubiquitin-Conjugating Enzymes / Q000235:genetics / Q000378:metabolism*; D000098843:Ubiquitin-Conjugating Enzyme UBC9
chemical_list: D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D025841:Small Ubiquitin-Related Modifier Proteins; D044763:Ubiquitin-Conjugating Enzymes; D048728:MAP Kinase Kinase Kinase 1; D000098843:Ubiquitin-Conjugating Enzyme UBC9
references: Cell. 2002 Jan 11;108(1):109-20
references: 11792325
references: Nat Methods. 2007 Mar;4(3):245-50
references: 17277783
references: EMBO J. 2002 Jun 3;21(11):2682-91
references: 12032081
references: Mol Cell Biol. 2003 Apr;23(8):2953-68
references: 12665592
references: Cell. 2003 Apr 4;113(1):127-37
references: 12679040
references: Mol Cell Biol. 2004 Mar;24(5):2052-62
references: 14966284
references: J Biol Chem. 2004 May 14;279(20):20999-1002
references: 15016812
references: Proc Natl Acad Sci U S A. 2004 Jun 8;101(23):8551-6
references: 15161980
references: J Biol Chem. 2004 Aug 6;279(32):33791-8
references: 15175327
references: Cell Stress Chaperones. 2004 Summer;9(2):214-20
references: 15497507
references: EMBO J. 1997 Sep 15;16(18):5509-19
references: 9312010
references: J Biol Chem. 1997 Oct 24;272(43):26799-802
references: 9341106
references: FEBS Lett. 1997 Nov 17;417(3):297-300
references: 9409737
references: Biochem Biophys Res Commun. 1999 Jan 27;254(3):693-8
references: 9920803
references: J Biol Chem. 1999 Apr 9;274(15):10618-24
references: 10187858
references: FEBS Lett. 1999 Apr 1;448(1):185-9
references: 10217437
references: Cancer Res. 2004 Nov 1;64(21):7846-51
references: 15520190
references: Biochim Biophys Acta. 2004 Nov 29;1695(1-3):113-31
references: 15571812
references: Mol Cell Proteomics. 2005 Jan;4(1):56-72
references: 15576338
references: J Biol Chem. 2005 Feb 11;280(6):5004-12
references: 15561718
references: J Biol Chem. 2005 Mar 18;280(11):9937-45
references: 15637059
references: Biochem Biophys Res Commun. 2005 Apr 22;329(4):1293-9
references: 15766567
references: Mol Cell. 2005 Apr 1;18(1):1-12
references: 15808504
references: Mol Cell Biol. 2005 Aug;25(16):6964-79
references: 16055710
references: Biochem Biophys Res Commun. 2005 Dec 2;337(4):1308-18
references: 16236267
references: Proc Natl Acad Sci U S A. 2006 Jan 3;103(1):45-50
references: 16371476
references: EMBO J. 1999 Nov 15;18(22):6455-61
references: 10562557
references: EMBO J. 1999 Nov 15;18(22):6462-71
references: 10562558
references: J Biol Chem. 2000 Mar 3;275(9):6252-8
references: 10692421
references: Proc Natl Acad Sci U S A. 2000 Dec 19;97(26):14145-50
references: 11121022
references: J Biol Chem. 2001 Apr 20;276(16):12654-9
references: 11124955
references: Mol Cell. 2001 Sep;8(3):713-8
references: 11583632
references: Cell. 2001 Oct 5;107(1):5-8
references: 11595179
references: Mol Cell. 2006 Jun 23;22(6):783-94
references: 16793547
references: EMBO Rep. 2006 Aug;7(8):838-44
references: 16845370
references: Proc Natl Acad Sci U S A. 2002 Mar 5;99(5):2872-7
references: 11867732