Abstract
Our previous studies demonstrated that JWA, a novel retinoic acids responsive and cytoskeleton related gene, is associated with cell differentiation and apoptosis. In the present study, to elucidate if the JWA is a novel kind of microtubule-associated proteins (MAPs) and functionally link to microtubule, we first successfully identified JWA from the physically purified MAPs complex of rat brain tissues. The results of co-immunoprecipitation, gene transfection and immunofluorescence microscopy assays from HBE and NIH3T3 cells provide strong evidence for a linkage between JWA and β-tubulin. In general, JWA is stably binding to β-tubulin whenever microtubule is polymerized or not, and it may be critical to the mitosis process. In addition, by use of the antisense oligonucleotides technique, we also showed that JWA is a negative modulator on intracellular amino acids in PC 12 cells. Further analysis indicated that JWA selectively regulates both taurine, an inhibitory amino acid, and glutamate, an excitatory amino acid. In conclusion, JWA is not only structurally associated, but also a novel functional MAP.
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Bifulco, M., Laezza, C., Stingo, S. et al., 2′, 3′-Cyclic nucleotide 3′ -phosphodiesterase: a membrane-bound, microtubule associated protein and membrane anchor for tubulin, Proc. Natl. Acad. Sci., 2002, 99: 1807–1812.
Bernier-Valentin, F., Aunis, D., Rousset, B., Evidence for tubulin-binding sites on cellular membranes: plasma membranes, mitochondrial membranes, and secretory granule membranes, J. Cell Biol., 1983, 97: 209–216.
Smith, D. S., Jarlfors, U., Cayer, M. L., Structural cross-bridges between microtubules and mitochondria in central axons of an insect (Periplaneta americana), J. Cell Sci., 1977, 27: 255–272.
Heggeness, M. H., Simon, M., Singer, S. J., Association of mitochondria with microtubules in cultured cells, Proc. Natl. Acad. Sci., 1978, 75: 3863–3866.
Gray, E. G., Presynaptic microtubules and their association with synaptic vesicles, Proc. R. Soc. Lond. B. Biol. Sci., 1975, 190: 367–372.
Vogl, A. W., Spatially dynamic intercellular adhesion junction is coupled to a microtubule-based motility system: evidence from anin vitro binding assay, Cell Motil. Cytoskeleton, 1996, 34: 1–12.
Mithieux, G., Audebet, C., Rousset, B., Association of purified thyroid lysosomes to reconstituted microtubules, Biochim. Biophys. Acta, 1988, 969: 121–130.
Dentier, W. L., Microtubule-membrane interactions in cilia and flagella, Int. Rev. Cytol., 1981, 72: 1–47.
Hardham, A. R., Gunning, B. E., Structure of cortical microtubule arrays in plant cells, J. Cell Biol., 1978, 77: 14–34.
Xia, W., Zhou, J. W., Cao, H. X. et al., Relationship between structure and functionof JWA in modulation of cell differentiation, Chinese Science Bulletin, 2001, 46(24): 2063–2067.
Cao, H. X., Xia, W., Shen, Q. et al., Role ofJWA in acute promyelocytic leukemia cell differentiation and apoptosis triggered by retinoic acid, 12-tetradecanoylphorbol-13-acetate and arsentic trioxide, Chinese Science Bulletin, 2002, 47(10): 834–838.
Zhou, J. W., Di, Y. P., Zhao, Y. H. et al., A novel cytoskeleton associate gene-clonging, identification, sequencing, regulation of expression and tissue distribution ofJWA (in Chinese), in Investigation on Cell Modulation: Signal Transduction, Apoptosis and Gene Expression (eds. Ye, X. S., Shen, B. F., Tang, X. F. et al.), Beijing: Military Medical Sciences Press, 1999, 110–119.
Ann, D. K., Wu, M. M., Wu, R. et al., Retinol-regulated gene expression in human tracheobronchial epithelial cells, J. Biol. Chem., 1988, 263: 3546–3549.
Nogales, E., Medrano, F. J., Diakun, G. P. et al., The effect of temperature on the structure of vinblastine-induced polymers of purified tubulin: detection of a reversible conformational change, J. Mol. Biol., 1995, 254: 416–430.
Bhattacharyya, B., Wolff, J., Polymerisation of membrane tubulin, Nature, 1976, 264: 567–577.
Abdul-Ghani, M., Gougeon, P. Y., Prosser, D. C. et al., PRA isoforms are targeted to distinct membrane compartments, J. Biol. Chem., 2001, 276: 6225–6233.
Chavrier, P., Vingron, M., Sander, C. et al., Molecular cloning of YPTl/SEC4-related cDNAs from an epithelial cell line, Mol. Cell Biol., 1990, 10: 6578–6585.
Matteoli, M., Takei, K., Cameron, R. et al., Association of Rab3A with synaptic vesicles at late stages of the secretory pathway, J. Cell Biol., 1991, 115: 625–633.
Elferink, L. A., Anzai, K., Seheller, R. H., Rabl5, a novel low molecular weight GTP-binding protein specifically expressed in rat brain, J. Biol. Chem., 1992, 267: 5768–5775.
Ngsee, J. K., Elferink, L. A., Seheller, R. H., A family of ras-like GTP-binding proteins expressed in electromotor neurons, J. Biol. Chem., 1991, 266: 2675–2680.
Fischer, von Mollard, G., Mignery, G. A., Baumert, M. et al., Rab3 is a small GTP-binding protein exclusively localized to synaptic vesicles, Proc. Natl. Acad. Sci., 1990, 87: 1988–1992.
Lazar, T., Gotte, M., Gallwitz, D., Vesicular transport: how many Ypt/Rab-GTPases make an eukaryotic cell, Trends Biochem. Sci., 1997, 22: 468–472.
Novick, P., Zerial, M., The diversity of Rab proteins in vesicle transport, Curr. Opin. Cell Biol., 1997, 9: 496–504.
Martinez, O., Goud, B., Rab proteins, Biochim. Biophys. Acta, 1998, 1404: 101–112.
Chen, W. Q., Jin, H., Nguyen, M. et al., Role of taurine in regulation of intracellular calcium level and neuroprotective function in cultured neurons, J. Neurosci. Res., 2001, 66: 612–619.
Saransaari, P., Oja, S. S., Taurine and neural cell damage, Amino Acids, 2000, 19: 509–526.
Lin, C. L., Orlov, I., Ruggiero, A. M. et al., Modulation of the neuronal glutamate transporter EAAC 1 by the interacting protein GTRAP3-18, Nature, 2001, 410: 84–88.
Butchbach, M. E., Guo, H., Lin, C. L., Methyl-beta-cyclodextrin but not retinoic acid reduces EAAT3-mediated glutamate uptake and increases GTRAP3-18 expression, J. Neurochem., 2003, 84: 891–894
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Li, A., Li, A., Mao, W. et al. JWA, a novel microtubule-associated protein, regulates homeostasis of intracellular amino acids in PC12 cells. Chin.Sci.Bull. 48, 1828–1834 (2003). https://doi.org/10.1007/BF03184063
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DOI: https://doi.org/10.1007/BF03184063