The small ubiquitin-like modifier protein (SUMO) regulates transcriptional activity and the translocation of proteins across the nuclear membrane1. The identification of SUMO substrates outside the nucleus is progressing2 but little is yet known about the wider cellular role of protein SUMOylation. Here we report that in rat hippocampal neurons multiple SUMOylation targets are present at synapses and we show that the kainate receptor subunit GluR6 is a SUMO substrate. SUMOylation of GluR6 regulates endocytosis of the kainate receptor and modifies synaptic transmission. GluR6 exhibits low levels of SUMOylation under resting conditions and is rapidly SUMOylated in response to a kainate but not an N-methyl-D-aspartate (NMDA) treatment. Reducing GluR6 SUMOylation using the SUMO-specific isopeptidase SENP-1 prevents kainate-evoked endocytosis of the kainate receptor. Furthermore, a mutated non-SUMOylatable form of GluR6 is not endocytosed in response to kainate in COS-7 cells. Consistent with this, electrophysiological recordings in hippocampal slices demonstrate that kainate-receptor-mediated excitatory postsynaptic currents are decreased by SUMOylation and enhanced by deSUMOylation. These data reveal a previously unsuspected role for SUMO in the regulation of synaptic function.
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We acknowledge the Wellcome Trust (J.M.H.), the MRC (J.R.M. and J.M.H.) and the EU (J.M.H. from a GRIPPANT contract) for financial support. We thank G. Hodgkinson for validation electrophysiology experiments on recombinant GluR6 in HEK cells, and S. Correa for some preliminary immuno cytochemistry experiments. We also thank M. Fleck for anti-GluR6 antibody, S. Goldstein for the SENP-1 constructs, C. Mulle for the pcDNA3-myc-GluR6 and H. Saitoh for the bacterial SUMOylation system. We are grateful to M. Ashby, G. Banting, Z. Bashir, T. Bouschet, G. Collingridge, J. Hanley, J. Isaac, F. Jaskolski, A. Randall and D. Stephens for commenting on the manuscript.
Author Contributions S.M. and A. N. are co-first authors; J.R.M. and J.M.H. are co-last authors. S.M. performed surface expression, biochemistry and cell imaging assays in cell culture. A.N. made the original observation that GluR6a is a SUMOylation substrate, performed biochemistry on brain extracts and performed all molecular biological experiments. J.R.M. performed the electrophysiology experiments and co-wrote the manuscript. J.M.H. provided team leadership, project management and wrote the manuscript. All authors contributed to hypothesis development, experimental design and data interpretation.
Reprints and permissions information is available at www.nature.com/reprints. The authors declare no competing financial interests.
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Martin, S., Nishimune, A., Mellor, J. et al. SUMOylation regulates kainate-receptor-mediated synaptic transmission. Nature 447, 321–325 (2007). https://doi.org/10.1038/nature05736
Cellular and Molecular Life Sciences (2022)
Molecular Neurodegeneration (2019)
Scientific Reports (2019)
Scientific Reports (2019)
Neurochemical Research (2019)