title: In Vitro Studies Reveal a Sequential Mode of Chain Processing by the Yeast SUMO (Small Ubiquitin-related Modifier)-specific Protease Ulp2.
abstract: Sumoylation is a post-translational modification essential in most eukaryotes that regulates stability, localization, activity, or interaction of a multitude of proteins. It is a reversible process wherein counteracting ligases and proteases, respectively, mediate the conjugation and deconjugation of SUMO molecules to/from target proteins. Apart from attachment of single SUMO moieties to targets, formation of poly-SUMO chains occurs by the attachment of additional SUMO molecules to lysine residues in the N-terminal extensions of SUMO. In Saccharomyces cerevisiae there are apparently only two SUMO(Smt3)-specific proteases: Ulp1 and Ulp2. Ulp2 has been shown to be important for the control of poly-SUMO conjugates in cells and to dismantle SUMO chains in vitro, but the mechanism by which it acts remains to be elucidated. Applying an in vitro approach, we found that Ulp2 acts sequentially rather than stochastically, processing substrate-linked poly-SUMO chains from their distal ends down to two linked SUMO moieties. Furthermore, three linked SUMO units turned out to be the minimum length of a substrate-linked chain required for efficient binding to and processing by Ulp2. Our data suggest that Ulp2 disassembles SUMO chains by removing one SUMO moiety at a time from their ends (exo mechanism). Apparently, Ulp2 recognizes surfaces at or near the N terminus of the distal SUMO moiety, as attachments to this end significantly reduce cleavage efficiency. Our studies suggest that Ulp2 controls the dynamic range of SUMO chain lengths by trimming them from the distal ends.
authors.affiliation: From the Institute for Genetics, Biocenter, University of Cologne, D-50674 Cologne, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Eckhoff
authors.affiliation: From the Institute for Genetics, Biocenter, University of Cologne, D-50674 Cologne, Germany j.dohmen@uni-koeln.de.
authors.forename: R Jürgen
authors.initials: RJ
authors.lastname: Dohmen
mesh_terms: D003001:Cloning, Molecular; D010450:Endopeptidases / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D049452:Green Fluorescent Proteins / Q000378:metabolism; D016366:Open Reading Frames; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D025841:Small Ubiquitin-Related Modifier Proteins / Q000378:metabolism*
chemical_list: C099362:SMT3 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D025841:Small Ubiquitin-Related Modifier Proteins; D049452:Green Fluorescent Proteins; D010450:Endopeptidases; C405949:ULP2 protein, S cerevisiae
keywords: Saccharomyces cerevisiae; protease; protein degradation; small ubiquitin-like modifier (SUMO); sumoylation
references.citation: Mol Cell Proteomics. 2005 Mar;4(3):246-54
references.pmid: 15542864
references.citation: Mol Cell. 2013 Jun 6;50(5):625-36
references.pmid: 23644018
references.citation: J Biol Chem. 2004 Oct 29;279(44):45662-8
references.pmid: 15326169
references.citation: J Cell Biol. 2006 Sep 25;174(7):939-49
references.pmid: 17000875
references.citation: Biochem J. 2009 Jul 15;421(2):223-30
references.pmid: 19392659
references.citation: Mol Cell. 2008 Nov 7;32(3):301-5
references.pmid: 18995828
references.citation: Genetics. 2001 May;158(1):95-107
references.pmid: 11333221
references.citation: Mol Cell. 2014 Mar 20;53(6):1053-66
references.pmid: 24582501
references.citation: J Biol Chem. 2000 May 12;275(19):14102-6
references.pmid: 10799485
references.citation: Biochem J. 2010 Sep 1;430(2):335-44
references.pmid: 20590526
references.citation: Trends Biochem Sci. 2007 Jun;32(6):286-95
references.pmid: 17499995
references.citation: Nat Rev Mol Cell Biol. 2012 Dec;13(12):755-66
references.pmid: 23175280
references.citation: Mol Biol Evol. 2013 May;30(5):1172-87
references.pmid: 23393154
references.citation: Annu Rev Biochem. 2004;73:355-82
references.pmid: 15189146
references.citation: J Biol Chem. 2001 Sep 21;276(38):35368-74
references.pmid: 11451954
references.citation: Biochem J. 2010 Jun 1;428(2):133-45
references.pmid: 20462400
references.citation: EMBO Rep. 2011 Feb;12(2):142-8
references.pmid: 21252943
references.citation: IUBMB Life. 2008 Nov;60(11):734-42
references.pmid: 18666185
references.citation: Nat Struct Mol Biol. 2014 Oct;21(10):927-36
references.pmid: 25218447
references.citation: Trends Biochem Sci. 2012 Jan;37(1):23-31
references.pmid: 22018829
references.citation: Annu Rev Biochem. 1998;67:509-44
references.pmid: 9759496
references.citation: Nat Rev Mol Cell Biol. 2009 Aug;10(8):564-8
references.pmid: 19474794
references.citation: Nat Rev Mol Cell Biol. 2009 Aug;10(8):550-63
references.pmid: 19626045
references.citation: Mol Cell Biol. 2010 Aug;30(15):3734-6
references.pmid: 20566698
references.citation: Trends Cell Biol. 2007 Aug;17(8):370-6
references.pmid: 17768054
references.citation: Mol Cell. 2000 May;5(5):865-76
references.pmid: 10882122
references.citation: Mol Cell Biol. 2007 Oct;27(19):6948-61
references.pmid: 17664284
references.citation: EMBO J. 1997 Sep 15;16(18):5509-19
references.pmid: 9312010
references.citation: Sci Signal. 2009;2(72):ra24
references.pmid: 19471022
references.citation: J Cell Biol. 2013 Apr 1;201(1):145-63
references.pmid: 23547032
references.citation: J Biol Chem. 1997 Oct 10;272(41):25560-5
references.pmid: 9325273
references.citation: J Biol Chem. 2003 Nov 7;278(45):44113-20
references.pmid: 12941945
references.citation: PLoS One. 2014;9(6):e100692
references.pmid: 24971888
references.citation: Nature. 1999 Mar 18;398(6724):246-51
references.pmid: 10094048
references.citation: Sci Rep. 2013;3:1690
references.pmid: 23604351
references.citation: Methods Mol Biol. 2003;205:99-117
references.pmid: 12491882
references.citation: Mol Gen Genet. 2000 Jun;263(5):771-86
references.pmid: 10905345
references.citation: Biochem Soc Trans. 2010 Feb;38(Pt 1):46-9
references.pmid: 20074033
references.citation: J Biol Chem. 2008 Nov 14;283(46):32045-55
references.pmid: 18799455
references.citation: Biochem Res Int. 2012;2012:875148
references.pmid: 22811915
references.citation: J Biol Chem. 2011 Mar 25;286(12):10238-47
references.pmid: 21247896
references.citation: Mol Biol Cell. 2009 Apr;20(8):2196-206
references.pmid: 19225149
references.citation: Mol Cell Proteomics. 2008 Jan;7(1):132-44
references.pmid: 17938407
references.citation: EMBO Rep. 2012 Apr;13(4):339-46
references.pmid: 22370726
references.citation: Mol Cell Proteomics. 2006 Dec;5(12):2298-310
references.pmid: 17000644
references.citation: Mol Cell Biol. 2000 Apr;20(7):2367-77
references.pmid: 10713161
references.citation: Mol Cell Proteomics. 2005 Jan;4(1):73-83
references.pmid: 15596868
references.citation: Annu Rev Biochem. 2013;82:357-85
references.pmid: 23746258
references.citation: J Biol Chem. 2005 Feb 11;280(6):4102-10
references.pmid: 15590687
references.citation: Biochim Biophys Acta. 2014 Jan;1843(1):75-85
references.pmid: 24018209
references.citation: J Biol Chem. 2007 Nov 23;282(47):34167-75
references.pmid: 17728242
references.citation: J Biol Chem. 2004 Jul 30;279(31):32262-8
references.pmid: 15166219
references.citation: Biochim Biophys Acta. 2009 Mar;1792(3):155-62
references.pmid: 19162180
references.citation: J Biol Chem. 2002 Dec 6;277(49):47938-45
references.pmid: 12354763
references.citation: Cell. 2014 Jul 17;158(2):327-38
references.pmid: 24998930
references.citation: J Biol Chem. 2009 Mar 27;284(13):8223-7
references.pmid: 19008217
references.citation: Nature. 2009 Mar 26;458(7237):422-9
references.pmid: 19325621
references.citation: Mol Cell. 2002 Jun;9(6):1169-82
references.pmid: 12086615
references.citation: Genome Biol. 2014;15(7):422
references.pmid: 25315341
references.citation: Biochem Soc Trans. 2013 Apr;41(2):463-73
references.pmid: 23514139
references.citation: Trends Biochem Sci. 2008 May;33(5):201-8
references.pmid: 18403209
references.citation: Nature. 1997 Feb 20;385(6618):737-40
references.pmid: 9034192
references.citation: J Biol Chem. 2004 Oct 1;279(40):41346-51
references.pmid: 15292183
references.citation: Mol Cell Biol. 2010 Aug;30(15):3737-48
references.pmid: 20516210
references.citation: J Proteome Res. 2006 Apr;5(4):761-70
references.pmid: 16602682
references.citation: Ernst Schering Found Symp Proc. 2008;(1):1-16
references.pmid: 19202597