Abstract
Tyrosine phosphorylation can upregulate NMDA receptor activity during pathological and physiological alterations of synaptic strength. Here we describe downregulation of recombinant NR1/2A receptors by tyrosine dephosphorylation that requires agonist binding, but is independent of ion flux. The tyrosine residues involved in this new form of NMDA receptor modulation likely form a 'ring' adjacent to the last transmembrane domain. The downregulation was due to a reduction in the number of functional channels, and was blocked by co-expressing a dominant-negative μ2-subunit of the clathrin-adaptor protein AP-2. Our results provide a mechanism by which synaptic NMDA receptors can be modulated in a use-dependent manner even when the postsynaptic membrane is not sufficiently depolarized to relieve channel block by magnesium ions.
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References
Wang, Y., Small, D. L., Stanimirovic, D. B., Morley, P. & Durkin, J. P. AMPA receptor-mediated regulation of a Gi-protein in cortical neurons. Nature 389, 502–504 (1997).
Hayashi, T., Umemori, H., Mishina, M. & Yamamoto, T. The AMPA receptor interacts with and signals through the protein tyrosine kinase Lyn. Nature 397, 72–76 (1999).
Rodriguez-Moreno, A. & Lerma, J. Kainate receptor modulation of GABA release involves a metabotropic function. Neuron 20, 1211–1218 (1998).
Swope, S. L., Moss, S. I., Raymond, L. A. & Huganir, R. L. Regulation of ligand-gated ion channels by protein phosphorylation. Adv. Second Messenger Phosphoprotein Res. 33, 49–78 (1999).
Dingledine, R., Borges, K., Bowie, D. & Traynelis, S. F. The glutamate receptor ion channels. Pharmacol. Rev. 51, 7–61 (1999).
Wang, Y. T., Yu, X. M. & Salter, M. W. Ca2+-independent reduction of N-methyl-D-aspartate channel activity by protein tyrosine phosphatase. Proc. Natl. Acad. Sci. USA 93, 1721–1725 (1996).
Wang, Y. T. & Salter, M. W. Regulation of NMDA receptors by tyrosine kinases and phosphatases. Nature 369, 233–235 (1994).
Chen, C. & Leonard, J. P. Protein tyrosine kinase-mediated potentiation of currents from cloned NMDA receptors. J. Neurochem. 67, 194–200 (1996).
Köhr, G. & Seeburg, P. H. Subtype-specific regulation of recombinant NMDA receptor-channels by protein tyrosine kinases of the src family. J. Physiol. (Lond.) 492, 445–452 (1996).
Zheng, F., Gingrich, M. B., Traynelis S. F. & Conn, P. J. Tyrosine kinase potentiates NMDA receptor currents by reducing tonic zinc inhibition. Nat. Neurosci. 1, 185–191 (1998).
Yu, X. M., Askalan, R., Keil II, G. J. & Salter, M. W. NMDA channel regulation by channel-associated protein tyrosine kinase Src. Science 275, 674–678 (1997).
Lu, W.-Y. et al. G-protein coupled receptors act via protein kinase C and Src to regulate NMDA receptors. Nat. Neurosci. 2, 331–338 (1999).
Xiong, Z.-G. et al. Src potentiation of NMDA receptors in hippocampal and spinal neurons is not mediated by reducing zinc inhibition. J. Neurosci. 19, RC37, 1–6 (1999).
O'Dell, T. J., Kandel, E. R. & Grant, S. G. Long-term potentiation in the hippocampus is blocked by tyrosine kinase inhibitors. Nature 353, 558–560 (1991).
Lu, Y. M., Roder, J. C., Davidow, J. & Salter, M. W. Src activation in the induction of long-term potentiation in CA1 hippocampal neurons. Science 279, 1363–1367 (1998).
Colledge, M. & Froehner, S. C. Signals mediating ion channel clustering at the neuromuscular junction. Curr. Opin. Neurobiol. 8, 357–363 (1998).
Rosenmund, C. & Westbrook, G. L. Calcium-induced actin depolymerization reduces NMDA channel activity. Neuron 10, 805–814 (1993).
Ehlers, M. D., Zhang, S., Bernhardt, J. P. & Huganir, R. L. Inactivation of NMDA receptors by direct interaction of calmodulin with the NR1 subunit. Cell 84, 745–755 (1996).
Krupp, J. J., Vissel, B., Heinemann, S. F. & Westbrook, G. L. N-terminal domains in the NR2 subunit control desensitization of NMDA receptors. Neuron 20, 317–327 (1998).
Krupp, J. J., Vissel, B., Thomas, C. G., Heinemann, S. F. & Westbrook, G. L. Interactions of calmodulin and alpha-actinin with the NR1 subunit modulate Ca2+-dependent inactivation of NMDA receptors. J. Neurosci. 19, 1165–1174 (1999).
Tong, G. & Jahr, C. E. Regulation of glycine-insensitive desensitization of the NMDA receptor in outside-out patches. J. Neurophysiol. 72, 754–761 (1994).
Wang, J., Zhou, Y., Wen, H. & Levitan, I. B. Simultaneous binding of two protein kinases to a calcium-dependent potassium channel. J. Neurosci. 19, RC4, 1–7 (1999).
Swope, S. L. & Huganir, R. L. Binding of the nicotinic acetylcholine receptor to SH2 domains of Fyn and Fyk protein tyrosine kinases. J. Biol. Chem. 269, 29817–29824 (1994).
Holmes, T. C., Fadool, D. A., Ren, R. & Levitan, I. B. Association of Src tyrosine kinase with a human potassium channel mediated by SH3 domain. Science 274, 2089–2091 (1996).
Stern, P., Cik, M., Colquhoun, D. & Stephenson, F. A. Single channel properties of cloned NMDA receptors in a human cell line: comparison with results from Xenopus oocytes. J. Physiol. (Lond.) 476, 391–397 (1994).
Rosenmund, C., Feltz, A. & Westbrook, G. L. Synaptic NMDA receptor channels have a low open probability. J. Neurosci. 15, 2788–2795 (1995).
Marsh, M. & McMahon, H. T. The structural era of endocytosis. Science 285, 215–220 (1999).
Nesterov, A., Carter, R. E., Sorkina, T., Gill, G. N. & Sorkin, A. Inhibition of the receptor-binding function of clathrin adaptor protein AP-2 by dominant-negative mutant μ2 subunit and its effect on endocytosis. EMBO J. 18, 2489–2499 (1999).
Legendre, P., Rosenmund, C. & Westbrook, G. L. Inactivation of NMDA channels in cultured hippocampal neurons by intracellular calcium. J. Neurosci. 13, 674–684 (1993).
Yu, X. - M. & Salter, M. W. Gain control of NMDA-receptor currents by intracellular sodium. Nature 396, 469–474 (1998).
Tezuka, T., Umemori, H., Akiyama, T., Nakanishi, S. & Yamamoto, T. PSD-95 promotes Fyn-mediated tyrosine phosphorylation of the N-methyl-D-aspartate receptor subunit NR2A. Proc. Natl. Acad. Sci. USA 96, 435–440 (1999).
Lin, S.-Y. et al. Brain-derived neurotrophic factor enhances association of protein tyrosine phosphatase PTP1D with the NMDA receptor subunit NR2B in the cortical postsynaptic density. Mol. Brain Res. 70, 18–25 (1999).
Fadool, D. A. & Levitan, I. B. Modulation of olfactory bulb potassium current by tyrosine phosphorylation. J. Neurosci. 18, 6126–6137 (1998).
Moon, I. S., Apperson, M. L. & Kennedy, M. B. The major tyrosine-phosphorylated protein in the postsynaptic density fraction is N-methyl-D-aspartate receptor subunit 2B. Proc. Natl. Acad. Sci. USA 91, 3954–3958 (1994).
Lau, L.-F. & Huganir, R. L. Differential tyrosine phosphorylation of N-methyl-D-aspartate receptor subunits. J. Biol. Chem. 270, 20036–20041 (1995).
Dunak, A. W., Yasuda, R. P. & Wolfe, B. B. Developmental regulation of tyrosine phosphorylation of the NR2D NMDA glutamate receptor subunit in rat central nervous system. J. Neurochem. 71, 1926–1934 (1998).
Zhou, S. & Cantley, L. C. Recognition and specificity in protein tyrosine kinase-mediated signalling. Trends Biochem. Sci. 20, 470–475 (1995).
Shiratori, T. et al. Tyrosine phosphorylation controls internalization of CTLA-4 by regulating its interaction with clathrin-associated adaptor complex AP-2. Immunity 6, 583–589 (1997).
Man, H.-Y. et al. Regulation of AMPA receptor-mediated synaptic transmission by clathrin-dependent receptor internalization. Neuron 25, 649–662 (2000).
Jones, S. M., Howell, K. E., Henley, J. R., Cao, H. & McNiven, M. A. Role of dynamin in the formation of transport vesicles from the trans-Golgi network. Science 279, 573–577 (1998).
Kreitzer, G., Marmorstein, A., Okamoto, P., Vallee, R. & Rodriguez-Boulan, E. Kinesin and dynamin are required for post-Golgi transport of a plasma-membrane protein. Nat. Cell Biol. 2, 125–127 (2000).
Craven, S. E. & Bredt, D. S. Synaptic targeting of the postsynaptic density protein PSD-95 mediated by a tyrosine-based trafficking signal. J. Biol. Chem. 275, 20045–20051 (2000).
Gurd, J. W. & Bissoon, N. The N-methyl-D-aspartate receptor subunits NR2A and NR2B bind to the SH2 domains of phospholipase C-gamma. J. Neurochem. 69, 623–630 (1997).
Takagi, N. et al. The effect of transient global ischemia on the interaction of Src and Fyn with the N-methyl-D-aspartate receptor and postsynaptic densities: possible involvement of Src homology 2 domains. J. Cereb. Blood Flow Metab. 19, 880–888 (1999).
Osten, P. et al. Mutagenesis reveals a role for ABP/GRIP binding to GluR2 in synaptic surface accumulation of the AMPA receptor. Neuron 27, 313–325 (2000).
Lüscher, C. et al. Role of AMPA receptor cycling in synaptic transmission and plasticity. Neuron 24, 649–658 (1999).
Hollmann, M. et al. Zinc potentiates agonist-induced currents at certain splice variants of the NMDA receptor. Neuron 10, 943–954 (1993).
Ishii, T. et al. Molecular characterization of the family of the N-methyl-D-aspartate receptor subunits. J. Biol. Chem. 268, 2836–2843 (1993).
Horten, R. M., Hunt, H. D., Ho, S. N., Pullen, J. K. & Pease, L. R. Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension. Gene 77, 61–68 (1989).
Kobayashi, S. et al. Activation of pp60c-src depending on cell density in PC12 cells. J. Biol. Chem. 272, 16262–16267 (1997).
Acknowledgements
This work was supported by NIH grants MH46613 (G.L.W.), NS28709 (S.F.H.), the McKnight Foundation (S.F.H.), the John Adler Foundation (S.F.H.), fellowships from the Human Frontiers program (J.J.K. and B.V.), a Bundy Foundation award (B.V.) and a CJ Martin NHMRC of Australia award (B.V.). We thank C. Schulteis and K. Prasad for preliminary experiments, C. Thomas and L. Grimes for preparation of neuronal cultures, and G. Royle for encouragement.
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Vissel, B., Krupp, J., Heinemann, S. et al. A use-dependent tyrosine dephosphorylation of NMDA receptors is independent of ion flux. Nat Neurosci 4, 587–596 (2001). https://doi.org/10.1038/88404
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DOI: https://doi.org/10.1038/88404
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