The dynamics and mechanism of SUMO chain deconjugation by SUMO-specific proteases.
SUMOylation of proteins is a cyclic process that requires both conjugation and deconjugation of SUMO moieties. Besides modification by a single SUMO, SUMO chains have also been observed, yet the dynamics of SUMO conjugation/deconjugation remain poorly understood. Using a non-deconjugatable form of SUMO we demonstrate the underappreciated existence of SUMO chains in vivo, we highlight the importance of SUMO deconjugation, and we demonstrate the highly dynamic nature of the SUMO system. We show that SUMO-specific proteases (SENPs) play a crucial role in the dynamics of SUMO chains in vivo by constant deconjugation. Preventing deSUMOylation in Schizosaccharomyces pombe results in slow growth and a sensitivity to replication stress, highlighting the biological requirement for deSUMOylation dynamics. Furthermore, we present the mechanism of SUMO chain deconjugation by SENPs, which occurs via a stochastic mechanism, resulting in cleavage anywhere within a chain. Our results offer mechanistic insights into the workings of deSUMOylating proteases and highlight their importance in the homeostasis of (poly)SUMO-modified substrates.
Sanford-Burnham Medical Research Institute, La Jolla, California 92037, USA.
Miklós
M
Békés
John
J
Prudden
Tharan
T
Srikumar
Brian
B
Raught
Michael N
MN
Boddy
Guy S
GS
Salvesen
D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D010447:Peptide Hydrolases / Q000235:genetics / Q000378:metabolism*; D025842:SUMO-1 Protein / Q000235:genetics / Q000378:metabolism*; D012568:Schizosaccharomyces / Q000235:genetics / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000378:metabolism*; D058207:Sumoylation / Q000502:physiology*
D025842:SUMO-1 Protein; D029702:Schizosaccharomyces pombe Proteins; D010447:Peptide Hydrolases
Cell. 2002 Dec 27;111(7):1041-54
12507430
Nat Methods. 2006 Jul;3(7):533-9
16791211
EMBO J. 2005 Feb 9;24(3):439-51
15660128
Mol Cell. 2009 Mar 13;33(5):570-80
19285941
Nature. 2009 Sep 3;461(7260):114-9
19675569
Structure. 2004 Aug;12(8):1519-31
15296745
Cell. 1983 Aug;34(1):11-2
6309404
Mol Cell Proteomics. 2008 Nov;7(11):2107-22
18565875
Cell Death Differ. 2010 Jan;17(1):25-34
19373246
J Biol Chem. 2007 Sep 7;282(36):26217-24
17591783
Nat Protoc. 2010 May;5(5):873-82
20431533
J Cell Biol. 2010 Mar 8;188(5):681-92
20212317
Mol Cell. 2008 Nov 7;32(3):301-5
18995828
Cell. 2006 Mar 24;124(6):1197-208
16564012
Mol Cell Proteomics. 2009 Dec;8(12):2664-75
19721078
Nat Rev Mol Cell Biol. 2007 Dec;8(12):947-56
18000527
Trends Cell Biol. 2007 Aug;17(8):370-6
17768054
Sci Signal. 2009 May 26;2(72):ra24
19471022
Mol Cell Biol. 2006 Jun;26(12):4489-98
16738315
J Biol Chem. 2008 Nov 14;283(46):32045-55
18799455
Nat Chem Biol. 2010 Oct;6(10):750-7
20802491
Mol Cell Biol. 2010 Sep;30(18):4452-62
20647537
Cell Death Differ. 2010 Jan;17(1):14-24
19893571
Thromb Haemost. 1995 Jul;74(1):139-48
8578447
Biochem J. 2010 Sep 1;430(2):335-44
20590526
Cell. 2008 Aug 22;134(4):679-91
18724940
IUBMB Life. 2008 Nov;60(11):734-42
18666185
Mol Cell Biol. 1999 Dec;19(12):8660-72
10567589
Anal Biochem. 2004 Aug 1;331(1):204-6
15246018
EMBO J. 2007 Jun 6;26(11):2797-807
17491593
Mol Cell Proteomics. 2008 Jan;7(1):132-44
17938407
Annu Rev Biochem. 2009;78:363-97
19489724
Mol Cell. 2000 May;5(5):865-76
10882122
Nat Struct Mol Biol. 2006 Dec;13(12):1060-8
17099700
Proteomics. 2009 Feb;9(4):922-34
19180541
Nat Struct Mol Biol. 2010 Aug;17(8):939-47
20622874
EMBO J. 1997 Sep 15;16(18):5509-19
9312010
J Biol Chem. 2003 Nov 7;278(45):44113-20
12941945
EMBO Rep. 2009 May;10(5):466-73
19373254
Proc Natl Acad Sci U S A. 2005 Oct 25;102(43):15587-92
16227443
Nature. 2010 Feb 18;463(7283):906-12
20164921
Mol Cell Proteomics. 2006 Dec;5(12):2298-310
17000644
EMBO J. 2007 Sep 19;26(18):4089-101
17762865
Biochem Biophys Res Commun. 2005 Nov 18;337(2):517-20
16198310
Nature. 2005 Jun 2;435(7042):687-92
15931224
Mol Cell. 2009 Jan 16;33(1):124-35
19150434
Trends Biochem Sci. 2003 Jun;28(6):321-8
12826404
J Biol Chem. 2001 Sep 21;276(38):35368-74
11451954
Nature. 1999 Mar 18;398(6724):246-51
10094048
Biochem J. 2008 Jan 15;409(2):461-9
17916063
Mol Cell. 2010 Apr 23;38(2):191-201
20417598
Nat Cell Biol. 2008 May;10(5):547-55
18408733
Nat Cell Biol. 2008 May;10(5):538-46
18408734
Proteomics. 2010 Jan;10(2):254-65
20029837
Biochem Biophys Res Commun. 1967 Apr 20;27(2):157-62
6035483
Biochem J. 2006 Jul 15;397(2):279-88
16553580
J Mol Biol. 2007 Jun 8;369(3):608-18
17466333
Trends Biochem Sci. 2008 May;33(5):201-8
18403209
Mol Cell Biol. 2007 Oct;27(19):6948-61
17664284
Proteomics. 2010 Jan;10(2):337-42
19899083