Abstract
Voltage-gated Ca2+ channels (VGCCs) are important in regulating a variety of cellular functions in neurons. It remains poorly understood how VGCCs with different functions are sorted within neurons. Here we show that the t-complex testis-expressed 1 (tctex1) protein, a light-chain subunit of the dynein motor complex, interacts directly and selectively with N- and P/Q-type Ca2+ channels, but not L-type Ca2+ channels. The interaction is insensitive to Ca2+. Overexpression in hippocampal neurons of a channel fragment containing the binding domain for tctex1 significantly decreases the surface expression of endogenous N- and P/Q-type Ca2+ channels but not L-type Ca2+ channels, as determined by immunostaining. Furthermore, disruption of the tctex1–Ca2+ channel interaction significantly reduces the Ca2+ current density in hippocampal neurons. These results underscore the importance of the specific tctex1-channel interaction in determining sorting and trafficking of neuronal Ca2+ channels with different functionalities.
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References
Catterall, W.A. Structure and function of neuronal Ca2+ channels and their role in neurotransmitter release. Cell Calcium 24, 307–323 (1998).
Dunlap, K., Luebke, J.I. & Turner, T.J. Exocytotic Ca2+ channels in mammalian central neurons. Trends Neurosci. 18, 89–98 (1995).
Zhang, J.F. et al. Distinctive pharmacology and kinetics of cloned neuronal Ca2+ channels and their possible counterparts in mammalian CNS neurons. Neuropharmacol. 32, 1075–1088 (1993).
Wheeler, D.B., Randall, A. & Tsien, R.W. Roles of N-type and Q-type Ca2+ channels in supporting hippocampal synaptic transmission. Science 264, 107–111 (1994).
West, A.E., Griffith, E.C. & Greenberg, M.E. Regulation of transcription factors by neuronal activity. Nat. Rev. Neurosci. 3, 921–931 (2002).
Westenbroek, R.E., Ahlijanian, M.K. & Catterall, W.A. Clustering of L-type Ca2+ channels at the base of major dendrites in hippocampal pyramidal neurons. Nature 347, 281–284 (1990).
Westenbroek, R.E. et al. Biochemical properties and subcellular distribution of an N-type calcium channel alpha 1 subunit. Neuron 9, 1099–1115 (1992).
Westenbroek, R.E. et al. Immunochemical identification and subcellular distribution of the α1A subunits of brain calcium channels. J. Neurosci. 15, 6403–6418 (1995).
Cohen-Cory, S. The developing synapse: construction and modulation of synaptic structures and circuits. Science 298, 770–776 (2002).
Garner, C.C., Zhai, R.G., Gundelfinger, E.D. & Ziv, N.E. Molecular mechanisms of CNS synaptogenesis. Trends Neurosci. 25, 243–251 (2002).
Sheng, M. Molecular organization of the postsynaptic specialization. Proc. Natl. Acad. Sci. USA 98, 7058–7061 (2001).
Nishimune, H., Sanes, J.R. & Carlson, S.S. A synaptic laminin-calcium channel interaction organizes active zones in motor nerve terminals. Nature 432, 580–587 (2004).
Catterall, W.A. Interactions of presynaptic Ca2+ channels and snare proteins in neurotransmitter release. Ann. NY Acad. Sci. 868, 144–159 (1999).
Chen, Y. et al. Formation of an endophilin-Ca2+ channel complex is critical for clathrin-mediated synaptic vesicle endocytosis. Cell 115, 37–48 (2003).
Herlitze, S. et al. Targeting mechanisms of high voltage-activated Ca2+ channels. J. Bioenerg. Biomembr. 35, 621–637 (2003).
Vallee, R.B., Williams, J.C., Varma, D. & Barnhart, L.E. Dynein: an ancient motor protein involved in multiple modes of transport. J. Neurobiol. 58, 189–200 (2004).
Hirokawa, N. Kinesin and dynein superfamily proteins and the mechanism of organelle transport. Science 279, 519–526 (1998).
Karcher, R.L., Deacon, S.W. & Gelfand, V.I. Motor-cargo interactions: the key to transport specificity. Trends Cell Biol. 12, 21–27 (2002).
Vale, R.D. The molecular motor toolbox for intracellular transport. Cell 112, 467–480 (2003).
Setou, M., Nakagawa, T., Seog, D.H. & Hirokawa, N. Kinesin superfamily motor protein KIF17 and mLin-10 in NMDA receptor-containing vesicle transport. Science 288, 11796–11802 (2000).
Tai, A.W., Chuang, J.Z., Bode, C., Wolfrum, U. & Sung, C.H. Rhodopsin's carboxy-terminal cytoplasmic tail acts as a membrane receptor for cytoplasmic dynein by binding to the dynein light chain Tctex-1. Cell 97, 877–887 (1999).
Maeno-Hikichi, Y. et al. A PKCε-ENH-channel complex specifically modulates N-type Ca2+ channels. Nat. Neurosci. 6, 468–475 (2003).
King, S.M. et al. The mouse t-complex-encoded protein Tctex-1 is a light chain of brain cytoplasmic dynein. J. Biol. Chem. 271, 32281–32287 (1996).
Kamal, A. & Goldstein, L.S. Principles of cargo attachment to cytoplasmic motor proteins. Curr. Opin. Cell Biol. 14, 63–68 (2002).
Llinas, R., Sugimori, M. & Silver, R.B. Microdomains of high calcium concentration in a presynaptic terminal. Science 256, 677–679 (1992).
Williams, M.E. et al. Structure and functional expression of an ω-conotoxin-sensitive human N-type calcium channel. Science 257, 389–395 (1992).
Gordon, G.W., Berry, G., Liang, X.H., Levine, B. & Herman, B. Quantitative fluorescence resonance energy transfer measurements using fluorescence microscopy. Biophys. J. 74, 2702–2713 (1998).
Jiang, X. & Sorkin, A. Coordinated traffic of Grb2 and Ras during epidermal growth factor receptor endocytosis visualized in living cells. Mol. Biol. Cell. 13, 1522–1535 (2002).
Siegel, R.M. et al. Measurement of molecular interactions in living cells by fluorescence resonance energy transfer between variants of the green fluorescent protein. Sci. STKE 2000, PL1 (2000).
Erickson, M.G., Alseikhan, B.A., Peterson, B.Z. & Yue, D.T. Preassociation of calmodulin with voltage-gated Ca2+ channels revealed by FRET in single living cells. Neuron. 31, 973–985 (2001).
Dotti, C., Sullivan, C. & Banker, G. The establishment of polarity by hippocampal neurons in culture. J. Neurosci. 8, 1454–1468 (1988).
Bartlett, W. & Banker, G. An electron microscopic study of the development of axons and dendrites by hippocampal neurons in culture. II. Synaptic relationships. J. Neurosci. 4, 1954–1965 (1984).
Pravettoni, E. et al. Different localizations and functions of L-type and N-type calcium channels during development of hippocampal neurons. Dev. Biol. 227, 581–594 (2000).
Jones, O.T. et al. N-type calcium channels in the developing rat hippocampus: subunit, complex, and regional expression. J. Neurosci. 17, 6152–6164 (1997).
Bahls, F.H. et al. Contact-dependent regulation of N-type calcium channel subunits during synaptogenesis. J. Neurobiol. 35, 198–208 (1998).
Maximov, A. & Bezprozvanny, I. Synaptic targeting of N-type calcium channels in hippocampal neurons. J. Neurosci. 22, 6939–6952 (2002).
King, S.M. et al. Cytoplasmic dynein contains a family of differentially expressed light chains. Biochem. 37, 15033–15041 (1998).
Fuhrmann, J.C. et al. Gephyrin interacts with dynein light chains 1 and 2, components of motor protein complexes. J. Neurosci. 22, 5393–5402 (2002).
Schwarzer, C., Barnikol-Watanabe, S., Thinnes, F.P. & Hilschmann, N. Voltage-dependent anion-selective channel (VDAC) interacts with the dynein light chain Tctex1 and the heat-shock protein PBP74. Int. J. Biochem. Cell Biol. 34, 1059–1070 (2002).
Nagano, F. et al. Interaction of Doc2 with tctex-1, a light chain of cytoplasmic dynein. Implication in dynein-dependent vesicle transport. J. Biol. Chem. 273, 30065–30068 (1998).
Yano, H. et al. Association of Trk neurotrophin receptors with components of the cytoplasmic dynein motor. J. Neurosci. 21, RC125 (2001).
Campbell, K.S., Cooper, S., Dessing, M., Yates, S. & Buder, A. Interaction of p59fyn kinase with the dynein light chain, Tctex-1, and colocalization during cytokinesis. J. Immunol. 161, 1728–1737 (1998).
Mok, Y.K., Lo, K.W. & Zhang, M. Structure of Tctex-1 and its interaction with cytoplasmic dynein intermediate chain. J. Biol. Chem. 276, 14067–14074 (2001).
Susalka, S.J., Hancock, W.O. & Pfister, K.K. Distinct cytoplasmic dynein complexes are transported by different mechanisms in axons. Biochim. Biophys. Acta. 1496, 76–88 (2000).
Tai, A.W., Chuang, J.Z. & Sung, C.H. Localization of Tctex-1, a cytoplasmic dynein light chain, to the Golgi apparatus and evidence for dynein complex heterogeneity. J. Biol. Chem. 273, 19639–19649 (1998).
Chuang, J.Z., Milner, T.A. & Sung, C.H. Subunit heterogeneity of cytoplasmic dynein: Differential expression of 14 kDa dynein light chains in rat hippocampus. J. Neurosci. 21, 5501–5512 (2001).
Augustine, G.J. How does calcium trigger neurotransmitter release? Curr. Opin. Neurobiol. 11, 320–326 (2001).
Rizo, J. & Sudhof, T.C. Snares and Munc18 in synaptic vesicle fusion. Nat. Rev. Neurosci. 3, 641–653 (2002).
Gundelfinger, E.D., Kessels, M.M. & Qualmann, B. Temporal and spatial coordination of exocytosis and endocytosis. Nat. Rev. Mol. Cell Biol. 4, 127–139 (2003).
Verhage, M. et al. DOC2 proteins in rat brain: complementary distribution and proposed function as vesicular adapter proteins in early stages of secretion. Neuron 18, 453–461 (1997).
Acknowledgements
We thank R. Pittman for the use of a Nikon Eclipse TE2000 inverted microscope; K. Campbell and C.H. Sung for anti-tctex1 antibody; M. Maronski and M. Dichter for help with hippocampal neuron cultures; P. Baas, K. Pfister and J. Meinkoth for helpful discussions and comments; and J. Field for initial help with yeast two-hybrid screening. This work was supported by grants from the US National Institutes of Health (J.F.Z. & W.A.S.), the American Heart Association (J.F.Z.) and the National Alliance for Research on Schizophrenia and Depression (Y.M.H.).
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Lai, M., Wang, F., Rohan, J. et al. A tctex1-Ca2+ channel complex for selective surface expression of Ca2+ channels in neurons. Nat Neurosci 8, 435–442 (2005). https://doi.org/10.1038/nn1418
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DOI: https://doi.org/10.1038/nn1418
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