TY - JOUR
T1 - A molecular basis for phosphorylation-dependent SUMO conjugation by the E2 UBC9
AU - Mohideen, Firaz
AU - Capili, Allan D.
AU - Bilimoria, Parizad M.
AU - Yamada, Tomoko
AU - Bonni, Azad
AU - Lima, Christopher D.
N1 - Funding Information:
We thank the NMR staff, particularly K. Dutta, at the New York Structural Biology Center (NYSBC) for assistance in NMR data collection and processing. We also thank A. Shalizi and A.Yunus for helpful discussions, and N. Arango for help with 293T transfection assays. NMR resources at NYSBC are supported by the US National Institutes of Health (NIH) grant P41 GM66354. The work was supported in part by NIH grants GM075695 (A.D.C.), NS041021 (A.B.) and GM065872 (C.D.L.).
PY - 2009/9
Y1 - 2009/9
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=69949179260&partnerID=8YFLogxK
U2 - 10.1038/nsmb.1648
DO - 10.1038/nsmb.1648
M3 - Article
C2 - 19684601
AN - SCOPUS:69949179260
SN - 1545-9993
VL - 16
SP - 945
EP - 952
JO - Nature Structural and Molecular Biology
JF - Nature Structural and Molecular Biology
IS - 9
ER -