A molecular basis for phosphorylation-dependent SUMO conjugation by the E2 UBC9.
Phosphorylation and small ubiquitin-like modifier (SUMO) conjugation contribute to the spatial and temporal regulation of substrates containing phosphorylation-dependent SUMO consensus motifs (PDSMs). Myocyte-enhancement factor 2 (MEF2) is a transcription factor and PDSM substrate whose modification by SUMO drives postsynaptic dendritic differentiation. NMR analysis revealed that the human SUMO E2 interacted with model substrates for phosphorylated and nonphosphorylated MEF2 in similar extended conformations. Mutational and biochemical analysis identified a basic E2 surface that enhanced SUMO conjugation to phosphorylated PDSM substrates MEF2 and heat-shock transcription factor 1 (HSF1), but not to nonphosphorylated MEF2 or HSF1, nor the non-PDSM substrate p53. Mutant ubiquitin-conjugating enzyme UBC9 isoforms defective in promoting SUMO conjugation to phosphorylated MEF2 in vitro and in vivo also impair postsynaptic differentiation in organotypic cerebellar slices. These data support an E2-dependent mechanism that underlies phosphorylation-dependent SUMO conjugation in pathways that range from the heat-shock response to nuclear hormone signaling to brain development.
Mohideen
Firaz
F
Program in Structural Biology, Sloan-Kettering Institute, New York, New York, USA.
Capili
Allan D
AD
Bilimoria
Parizad M
PM
Yamada
Tomoko
T
Bonni
Azad
A
Lima
Christopher D
CD
D020816:Amino Acid Motifs; D000818:Animals; D055162:Biocatalysis; D002460:Cell Line; D002531:Cerebellum / Q000378:metabolism; D003712:Dendrites / Q000378:metabolism; D006801:Humans; D026161:MADS Domain Proteins / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D051381:Rats; D016415:Sequence Alignment; D025841:Small Ubiquitin-Related Modifier Proteins / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity; D044763:Ubiquitin-Conjugating Enzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000098843:Ubiquitin-Conjugating Enzyme UBC9
D026161:MADS Domain Proteins; D025841:Small Ubiquitin-Related Modifier Proteins; D044763:Ubiquitin-Conjugating Enzymes; D000098843:Ubiquitin-Conjugating Enzyme UBC9
J Biol Chem. 2002 Jun 14;277(24):21740-8
11877416
J Biomol NMR. 1994 Sep;4(5):603-14
22911360
Annu Rev Biochem. 2004;73:355-82
15189146
Nature. 2005 Jul 21;436(7049):428-33
15931174
Cell. 2002 Feb 8;108(3):345-56
11853669
J Biomol NMR. 1995 Nov;6(3):277-93
8520220
J Biol Chem. 2004 Jul 9;279(28):29551-7
15123625
J Neurosci. 1995 Aug;15(8):5727-38
7643214
Mol Cell. 2006 Sep 15;23(6):779-86
16973431
Nature. 2005 Jun 2;435(7042):687-92
15931224
Neuron. 2004 Jan 22;41(2):229-41
14741104
Mol Cell. 2008 Jun 6;30(5):539-40
18538649
Annu Rev Biochem. 1998;67:425-79
9759494
Methods Mol Biol. 2009;497:167-86
19107417
Neuron. 2003 Apr 10;38(1):33-46
12691662
Nat Rev Mol Cell Biol. 2007 Dec;8(12):947-56
18000527
J Biol Chem. 2006 Feb 17;281(7):4423-33
16356933
J Biol Chem. 2001 Jun 15;276(24):21664-9
11259410
BMC Biochem. 2006 Feb 14;7:5
16478538
EMBO J. 2005 Feb 9;24(3):439-51
15660128
Genes Cells. 2004 Nov;9(11):1017-29
15507114
Annu Rev Cell Dev Biol. 2006;22:159-80
16753028
Mol Cell. 2006 Nov 3;24(3):341-54
17081986
Mol Cell. 2005 Jul 1;19(1):123-33
15989970
J Cell Mol Med. 2006 Jan-Mar;10(1):132-44
16563226
Methods Enzymol. 2005;398:74-87
16275321
EMBO J. 2006 Nov 1;25(21):5083-93
17036045
Annu Rev Biophys Biomol Struct. 2007;36:131-50
17477837
Science. 2006 Feb 17;311(5763):1012-7
16484498
Proc Natl Acad Sci U S A. 2006 Jan 3;103(1):45-50
16371476
J Neurosci. 2007 Sep 12;27(37):10037-46
17855618
Mol Cell. 2008 Jun 6;30(5):610-9
18538659
Curr Opin Struct Biol. 2007 Dec;17(6):726-35
17919899
Cell Div. 2008 Jan 24;3:5
18218095
J Mol Biol. 2006 Mar 10;356(5):1065-72
16405996
Nat Struct Mol Biol. 2006 Jun;13(6):491-9
16732283
J Biomol NMR. 1999 Jan;13(1):89-90
10070749