Abstract
REGULATED fusion of secretory granules with the plasma membrane in secretory cells requires ATP, Ca2+ and cytosolic1-3 as well as membrane4 proteins. ATP-dependent steps in Ca2+-activated secretion from PC 12 cells require three cytosolic PEP proteins (priming in exocytosis proteins, PEP1–3)5,6, the identity of which will provide insights into the required ATP-using reactions. PEP3 was recently identified as phosphatidylinositol transfer protein (PtdlnsTP)6, and here we report that PEP1 consists of the type I phosphatidylinositol-4-phosphate 5-kinase (PtdlnsPSK). The roles of PEP3/PtdInsTP and PEP1/ PtdlnsPSK in sequential phosphoinositide recruitment and phosphorylation explains their synergistic activity in ATP-dependent priming. Moreover, inhibition of Ca2+-activated secretion by PtdIns(4,5)P2-specific antibodies and phospholipase C implies that 5-phosphorylated inositides play a novel, necessary role in the regulated secretory pathway. The results indicate that lipid kinase-mediated phosphorylation is an important basis for ATP use in the exocytotic pathway.
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References
Walent, J. H., Porter, B. W. & Martin, T. F. J. Cell 70, 765–775 (1992).
Morgan, A. & Burgoyne, R. D. Nature 355, 833–836 (1992).
Wu, Y. N., Vu, N.-D. & Wagner, P. D. Biochem. J. 285, 697–700 (1992).
Söllner, T. et al. Nature 362, 318–324 (1993).
Hay, J. C. & Martin, T. F. J. J. Cell Biol. 119, 139–151 (1992).
Hay, J. C. & Martin, T. F. J. Nature 366, 572–575 (1993).
Helmkamp, G. M. Jr, Harvey, M. S., Wirtz, K. W. A. & van Deenen, L. L. M. J. biol. Chem. 249, 6382–6389 (1974).
Cleves, A. E. et al. Cell 64, 789–800 (1991).
McGee, T. P., Skinner, H. B., Whitters, E. A., Henry, S. A. & Bankaitis, V. A. J. Cell Biol. 124, 273–287 (1994).
Thomas, G. M. H. et al. Cell 74, 919–928 (1993).
Carpenter, C. L. & Cantley, L. C. Biochemistry 29, 11147–11156 (1990).
Jenkins, G. H., Fisette, P. L. & Anderson, R. A. J. biol. Chem. 269, 11547–11554 (1994).
Scholz, G., Barritt, G. J. & Kwok, F. Eur. J. Biochem. 210, 461–466 (1992).
Bazenet, C. E., Ruano, A. R., Brockman, J. L. & Anderson, R. A. J. biol. Chem. 265, 18012–18022 (1990).
Schu, P. V. et al. Science 260, 88–91 (1993).
Fukami, K. et al. Nature 359, 150–152 (1992).
Eberhard, D. A., Cooper, C. L. Low, M. G. & Holz, R. W. Biochem. J. 268, 15–25 (1990).
Phillips, J. H. Biochem. J. 136, 579–587 (1973).
Tagaya, M., Wilson, D. W., Brunner, M., Arango, N. & Rothman, J. E. J. biol. Chem. 268, 2662–2666 (1993).
Harlan, J. E., Hajduk, P. J., Yoon, H. S. & Fesik, S. W. Nature 371, 168–170 (1994).
Perrin, D., Langley, O. K. & Aunis, D. Nature 326, 498–501 (1987).
Anderson, R. A. & Marchesi, V. T. Nature 318, 295–298 (1985).
Janmey, P. A. A. Rev. Physiol. 56, 169–191 (1994).
Bittner, M. A. & Holz, R. W. Cell. molec. Neurobiol. 13, 649–663 (1993).
Lawrence, G. W., Weller, U. & Dolly, J. O. Eur. J. Biochem. 222, 325–333 (1994).
Brown, H. A., Gutowski, S., Moomaw, C. R., Slaughter, C. & Sternweis, P. C. Cell 75, 1137–1144 (1993).
Liscovitch, M., Chalifa, V., Pertile, P., Chen, C.-S. & Cantley, L. C. J. biol. Chem. 269, 21403–21406 (1993).
Laemmli, U. K. Nature 227, 680–695 (1970).
Burnette, W. R. Analyt. Biochem. 112, 195–203 (1981).
Ausubel, F. M. et al. Current Protocols in Molecular Biology (Wiley, New York, 1994).
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Hay, J., Fisette, P., Jenkins, G. et al. ATP-dependent inositide phosphorylation required for Ca2+-activated secretion. Nature 374, 173–177 (1995). https://doi.org/10.1038/374173a0
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DOI: https://doi.org/10.1038/374173a0
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