title: A molecular basis for phosphorylation-dependent SUMO conjugation by the E2 UBC9.
abstract: 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.
authors.affiliation: Program in Structural Biology, Sloan-Kettering Institute, New York, New York, USA.
authors.forename: Firaz
authors.initials: F
authors.lastname: Mohideen
authors.forename: Allan D
authors.initials: AD
authors.lastname: Capili
authors.forename: Parizad M
authors.initials: PM
authors.lastname: Bilimoria
authors.forename: Tomoko
authors.initials: T
authors.lastname: Yamada
authors.forename: Azad
authors.initials: A
authors.lastname: Bonni
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Lima
mesh_terms: 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
chemical_list: D026161:MADS Domain Proteins; D025841:Small Ubiquitin-Related Modifier Proteins; D044763:Ubiquitin-Conjugating Enzymes; D000098843:Ubiquitin-Conjugating Enzyme UBC9
references.citation: J Biol Chem. 2002 Jun 14;277(24):21740-8
references.pmid: 11877416
references.citation: J Biomol NMR. 1994 Sep;4(5):603-14
references.pmid: 22911360
references.citation: Annu Rev Biochem. 2004;73:355-82
references.pmid: 15189146
references.citation: Nature. 2005 Jul 21;436(7049):428-33
references.pmid: 15931174
references.citation: Cell. 2002 Feb 8;108(3):345-56
references.pmid: 11853669
references.citation: J Biomol NMR. 1995 Nov;6(3):277-93
references.pmid: 8520220
references.citation: J Biol Chem. 2004 Jul 9;279(28):29551-7
references.pmid: 15123625
references.citation: J Neurosci. 1995 Aug;15(8):5727-38
references.pmid: 7643214
references.citation: Mol Cell. 2006 Sep 15;23(6):779-86
references.pmid: 16973431
references.citation: Nature. 2005 Jun 2;435(7042):687-92
references.pmid: 15931224
references.citation: Neuron. 2004 Jan 22;41(2):229-41
references.pmid: 14741104
references.citation: Mol Cell. 2008 Jun 6;30(5):539-40
references.pmid: 18538649
references.citation: Annu Rev Biochem. 1998;67:425-79
references.pmid: 9759494
references.citation: Methods Mol Biol. 2009;497:167-86
references.pmid: 19107417
references.citation: Neuron. 2003 Apr 10;38(1):33-46
references.pmid: 12691662
references.citation: Nat Rev Mol Cell Biol. 2007 Dec;8(12):947-56
references.pmid: 18000527
references.citation: J Biol Chem. 2006 Feb 17;281(7):4423-33
references.pmid: 16356933
references.citation: J Biol Chem. 2001 Jun 15;276(24):21664-9
references.pmid: 11259410
references.citation: BMC Biochem. 2006 Feb 14;7:5
references.pmid: 16478538
references.citation: EMBO J. 2005 Feb 9;24(3):439-51
references.pmid: 15660128
references.citation: Genes Cells. 2004 Nov;9(11):1017-29
references.pmid: 15507114
references.citation: Annu Rev Cell Dev Biol. 2006;22:159-80
references.pmid: 16753028
references.citation: Mol Cell. 2006 Nov 3;24(3):341-54
references.pmid: 17081986
references.citation: Mol Cell. 2005 Jul 1;19(1):123-33
references.pmid: 15989970
references.citation: J Cell Mol Med. 2006 Jan-Mar;10(1):132-44
references.pmid: 16563226
references.citation: Methods Enzymol. 2005;398:74-87
references.pmid: 16275321
references.citation: EMBO J. 2006 Nov 1;25(21):5083-93
references.pmid: 17036045
references.citation: Annu Rev Biophys Biomol Struct. 2007;36:131-50
references.pmid: 17477837
references.citation: Science. 2006 Feb 17;311(5763):1012-7
references.pmid: 16484498
references.citation: Proc Natl Acad Sci U S A. 2006 Jan 3;103(1):45-50
references.pmid: 16371476
references.citation: J Neurosci. 2007 Sep 12;27(37):10037-46
references.pmid: 17855618
references.citation: Mol Cell. 2008 Jun 6;30(5):610-9
references.pmid: 18538659
references.citation: Curr Opin Struct Biol. 2007 Dec;17(6):726-35
references.pmid: 17919899
references.citation: Cell Div. 2008 Jan 24;3:5
references.pmid: 18218095
references.citation: J Mol Biol. 2006 Mar 10;356(5):1065-72
references.pmid: 16405996
references.citation: Nat Struct Mol Biol. 2006 Jun;13(6):491-9
references.pmid: 16732283
references.citation: J Biomol NMR. 1999 Jan;13(1):89-90
references.pmid: 10070749