Tyrosine phosphorylation regulates the endocytosis and surface expression of GluN3A-containing NMDA receptors

Dhrubajyoti Chowdhury, Sonia Marco, Ian M. Brooks, Aitor Zandueta, Yijian Rao, Volker Haucke, John F. Wesseling, Steven Tavalin, Isabel Pérez-Otaño

Research output: Contribution to journalArticle

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Abstract

Selective control of receptor trafficking provides a mechanism for remodeling the receptor composition of excitatory synapses, and thus supports synaptic transmission, plasticity, and development. GluN3A (formerly NR3A) is a nonconventional member of the NMDA receptor (NMDAR) subunit family, which endows NMDAR channels with low calcium permeability and reduced magnesium sensitivity compared with NMDARs comprising only GluN1 and GluN2 subunits. Because of these special properties, GluN3A subunits act as a molecular brake to limit the plasticity and maturation of excitatory synapses, pointing toward GluN3A removal as a critical step in the development of neuronal circuitry. However, the molecular signals mediating GluN3A endocytic removal remain unclear. Here we define a novel endocytic motif (YWL), which is located within the cytoplasmic C-terminal tail of GluN3A and mediates its binding to the clathrin adaptor AP2. Alanine mutations within the GluN3A endocytic motif inhibited clathrin-dependent internalization and led to accumulation of GluN3A-containing NMDARs at the cell surface, whereas mimicking phosphorylation of the tyrosine residue promoted internalization and reduced cell-surface expression as shown by immunocytochemical and electrophysiological approaches in recombinant systems and rat neurons in primary culture. We further demonstrate that the tyrosine residue is phosphorylated by Src family kinases, and that Src-activation limits surface GluN3A expression in neurons. Together, our results identify a new molecular signal for GluN3A internalization that couples the functional surface expression of GluN3A-containing receptors to the phosphorylation state of GluN3A subunits, and provides a molecular framework for the regulation of NMDAR subunit composition with implications for synaptic plasticity and neurodevelopment.

Original languageEnglish (US)
Pages (from-to)4151-4164
Number of pages14
JournalJournal of Neuroscience
Volume33
Issue number9
DOIs
StatePublished - Feb 27 2013

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Endocytosis
N-Methyl-D-Aspartate Receptors
Tyrosine
Neuronal Plasticity
Phosphorylation
Synapses
Vesicular Transport Adaptor Proteins
Neurons
Clathrin
src-Family Kinases
Synaptic Transmission
Alanine
Magnesium
Tail
Permeability
Calcium
Mutation

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)

Cite this

Chowdhury, D., Marco, S., Brooks, I. M., Zandueta, A., Rao, Y., Haucke, V., ... Pérez-Otaño, I. (2013). Tyrosine phosphorylation regulates the endocytosis and surface expression of GluN3A-containing NMDA receptors. Journal of Neuroscience, 33(9), 4151-4164. https://doi.org/10.1523/JNEUROSCI.2721-12.2013

Tyrosine phosphorylation regulates the endocytosis and surface expression of GluN3A-containing NMDA receptors. / Chowdhury, Dhrubajyoti; Marco, Sonia; Brooks, Ian M.; Zandueta, Aitor; Rao, Yijian; Haucke, Volker; Wesseling, John F.; Tavalin, Steven; Pérez-Otaño, Isabel.

In: Journal of Neuroscience, Vol. 33, No. 9, 27.02.2013, p. 4151-4164.

Research output: Contribution to journalArticle

Chowdhury, D, Marco, S, Brooks, IM, Zandueta, A, Rao, Y, Haucke, V, Wesseling, JF, Tavalin, S & Pérez-Otaño, I 2013, 'Tyrosine phosphorylation regulates the endocytosis and surface expression of GluN3A-containing NMDA receptors', Journal of Neuroscience, vol. 33, no. 9, pp. 4151-4164. https://doi.org/10.1523/JNEUROSCI.2721-12.2013
Chowdhury, Dhrubajyoti ; Marco, Sonia ; Brooks, Ian M. ; Zandueta, Aitor ; Rao, Yijian ; Haucke, Volker ; Wesseling, John F. ; Tavalin, Steven ; Pérez-Otaño, Isabel. / Tyrosine phosphorylation regulates the endocytosis and surface expression of GluN3A-containing NMDA receptors. In: Journal of Neuroscience. 2013 ; Vol. 33, No. 9. pp. 4151-4164.
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AU - Rao, Yijian

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AU - Wesseling, John F.

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