Abstract
Controlling the number of functional γ-aminobutyric acid A (GABAA) receptors in neuronal membranes is a crucial factor for the efficacy of inhibitory neurotransmission. Here we describe the direct interaction of GABAA receptors with the ubiquitin-like protein Plic-1. Furthermore, Plic-1 is enriched at inhibitory synapses and is associated with subsynaptic membranes. Functionally, Plic-1 facilitates GABAA receptor cell surface expression without affecting the rate of receptor internalization. Plic-1 also enhances the stability of intracellular GABAA receptor subunits, increasing the number of receptors available for insertion into the plasma membrane. Our study identifies a previously unknown role for Plic-1, a modulation of GABAA receptor cell surface number, which suggests that Plic-1 facilitates accumulation of these receptors in dendritic membranes.
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References
Macdonald, R. L. & Olsen, R. W. GABAA receptor channels. Annu. Rev. Neurosci. 17, 569–602 (1994).
Rabow, L. E., Russek, S. J. & Farb, D. H. From ion currents to genomic analysis: recent advances in GABAA receptor research. Synapse 21, 189–274 (1995).
Davies, P. A., Hanna, M. C., Hales, T. G. & Kirkness, E. F. Insensitivity to anaesthetic agents conferred by a class of GABAA receptor subunit. Nature 385, 820–823 (1997).
Bonnert, T. P. et al. Theta, a novel γ-aminobutyric acid type A receptor subunit. Proc. Natl. Acad. Sci. USA 96, 9891–9896 (1999).
Unwin, N. Neurotransmitter action: opening of ligand-gated ion channels. Cell Suppl. 72, 31–41 (1993).
Essrich, C., Lorez, M., Benson, J. A., Fritschy, J. M. & Luscher, B. Postsynaptic clustering of major GABAA receptor subtypes requires the γ2 subunit and gephyrin. Nat. Neurosci. 1, 563–571 (1998).
Kneussel, M. et al. Loss of postsynaptic GABAA receptor clustering in gephyrin-deficient mice. J. Neurosci. 19, 9289–9297 (1999).
Feng, G. et al. Dual requirement for gephyrin in glycine receptor clustering and molybdoenzyme activity. Science 282, 1321–1324 (1998).
Kittler, J. T. et al. The subcellular distribution of GABARAP and its ability to interact with NSF suggest a role for this protein in the intracellular transport of GABAA receptors. Mol. Cell Neurosci. 18, 13–25 (2001).
Wang, H. B., Bedford, F. K., Brandon, N. J., Moss, S. J. & Olsen, R. W. GABAA-receptor-associated protein links GABAA receptors and the cytoskeleton. Nature 397, 69–72 (1999).
Wan Q, et al. Recruitment of functional GABAA receptors to postsynaptic domains by insulin. Nature 388, 686–689 (1997).
Nusser, Z., Hajos, N., Somogyi, P. & Mody, I. Increased number of synaptic GABAA receptors underlies potentiation at hippocampal inhibitory. Nature 395, 172–177 (1998).
Nusser, Z., Cull-Candy, S. & Farrant, M. Differences in synaptic GABAA receptor number underlie variation in GABA mini amplitude. Neuron 19, 697–709 (1997).
Moss, S. J. & Smart, T. G. Constructing inhibitory synapses. Nat. Rev. Neurosci. 2, 241–250 (2001).
Connolly, C. N. et al. Subcellular localization and endocytosis of homomeric γ2 subunit splice variants of γ-aminobutyric acid type A receptors. Mol. Cell Neurosci. 13, 259–271 (1999).
Connolly, C. N. et al. Cell surface stability of γ-aminobutyric acid type A receptors. Dependence on protein kinase C activity and subunit composition. J. Biol. Chem. 274, 36565–36572 (1999).
Kittler, J. T. et al. Constitutive endocytosis of GABAA receptors by an association with the adaptin AP2 complex modulates inhibitory synaptic currents in hippocampal neurons. J. Neurosci. 20, 7972–7977 (2000).
Kittler, J. T. et al. Analysis of GABAA receptor assembly in mammalian cell lines and hippocampal neurons using γ2 subunit green fluorescent protein chimeras. Mol. Cell Neurosci. 16, 440–452 (2000).
Marsh, M. & McMahon, H. T. The structural era of endocytosis. Science 285, 215–220 (1999).
Wu, A. L., Wang, J., Zheleznyak, A. & Brown, E. J. Ubiquitin-related proteins regulate interaction of vimentin intermediate filaments with the plasma membrane. Mol. Cell 4, 619–625 (1999).
Kleijnen, M. F. et al. The hPLIC proteins may provide a link between the ubiquitination machinery and the proteasome. Mol. Cell 6, 409–401 (2000).
Field., S. & Song, O. A novel genetic system to detect protein–protein interactions. Nature 340, 245–246 (1989).
Dong, H. et al. GRIP: a synaptic PDZ domain-containing protein that interacts with AMPA receptors. Nature 386, 279–284 (1997).
Funakoshi, M., Geley, S., Hunt, T., Nishimoto, T. & Kobayashi, H. Identification of XDRP1; a Xenopus protein related to yeast Dsk2p binds to the N-terminus of cyclin A and inhibits its degradation. EMBO J. 18, 5009–5018 (1999).
Jentsch, S. & Pyrowolakis, G. Ubiquitin and its kin: how close are the family ties? Trends Cell Biol. 10, 335–341 (2000).
Hofmann, K. & Bucher, P. The UBA domain: a sequence motif present in multiple enzyme classes of the ubiquitination pathway. Trends Biochem. Sci. 21, 172–173 (1996).
Hanley, J. G., Koulen, P., Bedford, F., Gordon-Weeks, P. R. & Moss, S. J. The protein MAP-1B links GABAC receptors to the cytoskeleton at retinal synapses. Nature 397, 66–90 (1999).
Smith, D. B. & Johnson, K. S. Single step purification of polypeptides expressed in E. Coli as fusions with glutathione S-transferase. Gene 67, 31–40 (1988).
Benke, D., Fritschy, J. M., Trzeciak, A., Bannwarth, W. & Mohler, H. Distribution, prevalence, and drug binding profile of gamma-aminobutyric acid type A receptor subtypes differing in the beta-subunit variant. J. Biol. Chem. 269, 27100–27107 (1994).
Wooltorton, J. R., Moss, S. J. & Smart, T. G. Pharmacological and physiological characterization of murine homomeric β3 GABAA receptors. Eur. J. Neurosci. 9, 2225–2235 (1997).
Connolly, C. N., McDonald, B. M., Krishek, B. J., Smart, T. G. & Moss, S. J. Assembly and cell surface expression of heteromeric and homomeric GABAA receptors. J. Biol. Chem. 271, 89–97 (1996).
Todd, A. J., Watt, C., Spike, R. C. & Sieghart, W. Co-localization of GABA, glycine, and their receptors at synapses in the rat spinal cord. J. Neurosci. 16, 974–982 (1996).
Gardiol, A., Racca, C. & Triller, A. Dendritic and postsynaptic protein synthetic machinery. J. Neurosci. 19, 168–179 (1999).
Noel, J. et al. Surface expression of AMPA receptors in hippocampal neurons is regulated by a NSF-dependent mechanism. Neuron 23, 365–376 (1999).
Williams, E. J. et al. Selective inhibition of growth factor-stimulated mitogenesis by a cell-permeable Grb2-binding peptide. J. Biol. Chem. 272, 22349–22354 (1997).
Taylor, P. M. et al. Identification of amino acid residues within GABAA receptor β subunits that mediate both homomeric and heteromeric receptor expression. J. Neurosci. 19, 6360–6371 (1999).
Taylor, P. M. et al. Identification of residues within GABAA receptor alpha subunits that mediate specific assembly with receptor beta subunits. J. Neurosci. 20, 1297–1306 (2000).
Garner, C. C., Nash, J. & Huganir, R. L. PDZ domains in synapse assembly and signaling. Trends Cell Biol. 10, 274–280 (2000).
Mah, A. L., Perry, G., Smith, M. A. & Monteiro, M. J. Identification of ubiquilin, a novel presenilin interactor that increases presenilin protein accumulation. J. Cell Biol. 151, 847–862 (2000).
Gorrie, G. H. et al. Assembly of GABAA receptors composed of α1 and β2 subunits in both cultured neurones and fibroblasts. J. Neurosci. 17, 6587–6588 (1997).
Colin, I., Rostaing, P., Augustin, A. & Triller, A. Localization of components of glycinergic synapses during rat spinal cord development. J. Comp. Neurol. 398, 359–372 (1998).
Triller, A., Cluzeaud, F., Pfeiffer, F., Betz, H. & Korn, H. Distribution of glycine receptors at central synapses: an immunoelectron microscopy study. Cell Biol. 101, 683–688 (1985).
Triller, A., Cluzeaud, F., Pfeiffer, F. & Korn, H. in Molecular Aspects of Neurobiology (eds. Levi Montalcini, R. et al.) 101–105 (Springer, Berlin, Heidelberg, 1986).
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This work was supported by the MRC, the Wellcome Trust and the Institut de Recherche sur la Moelle Epinière.
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Bedford, F., Kittler, J., Muller, E. et al. GABAA receptor cell surface number and subunit stability are regulated by the ubiquitin-like protein Plic-1. Nat Neurosci 4, 908–916 (2001). https://doi.org/10.1038/nn0901-908
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DOI: https://doi.org/10.1038/nn0901-908
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