Neurocrescin Is Specifically Cleaved after the Sequence DESD in a Caspase-3-Independent Manner
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1. A neurite outgrowth factor, neurocrescin (NC), which we previously identified from an extract of denervated skeletal muscle, was endoproteolytically processed in cell transfectants. In addition to the processing site identified in NC (DESD358/F) being similar to the optimal recognition sequence of group II caspases, DExD, cleavage site mutations confirmed the involvement of caspase(s) in NC processing.
2. However, both the recombinant NC and the synthetic octapeptide (YL DESDFG) were scarcely cleaved in vitro by caspase-3 or -7. Furthermore, transiently expressed NC was cleaved even in the caspase-3-deficient cell line, MCF-7 cells, and this efficiency was not altered by the transfectional expression of caspase-3.
3. Using the fluorescent substrate (Ac-DESD-AMC), the characteristic proteolytic activities, which cleaved it more effectively than caspase-3 and whose pH dependences were different from those of caspase-3, were endogenously identified in the muscle extract. These findings indicate the presence of proteolytic activities that are distinguishable from caspase-3.
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- Choi, E. K., Zaidi, N. F., Miller, J. S., Crowley, A. C., Merriam, D. E., Lilliehook, C., Buxbaum, J. D., and Wasco, W. (2001). Calsenilin is a substrate for caspase-3 that preferentially interacts with the familial Alzheimer’s disease-associated C-terminal fragment of presenilin 2. J. Biol. Chem. 276:19197–19204.Google Scholar
- Cosulich, S. C., Horiuchi, H., Zerial, M., Clarke, P. R., and Woodman, P. G.(1997). Cleavage of rabaptin-5 blocks endosome fusion during apoptosis. Embo. J. 16:6182–6191.Google Scholar
- Gavrieli, Y., Sherman, Y., and Ben-Sasson, S. A.(1992). Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J. Cell Biol. 119:493–501.Google Scholar
- Germain, M., Affar, E. B., D’Amours, D., Dixit, V. M., Salvesen, G. S., and Poirier, G. G.(1999). Cleavage of automodified poly(ADP-ribose) polymerase during apoptosis. Evidence for involvement of caspase-7. J. Biol. Chem. 274:28379–28384.Google Scholar
- Henderson, C. E., Huchet, M., and Changeux, J. P.(1981). Neurite outgrowth from embryonic chicken spinal neurons is promoted by media conditioned by muscle cells. Proc. Natl. Acad. Sci. USA 78:2625–2629.Google Scholar
- Hill, M. A., and Bennett, M. R.(1986). Motoneurone survival activity in extracts of denervated muscle reduced by prior stimulation of the muscle. Brain Res. 389:305–308.Google Scholar
- Ito, W., Ishiguro, H., and Kurosawa, Y.(1991). A general method for introducing a series of mutations into cloned DNA using the polymerase chain reaction. Gene 102:67–70.Google Scholar
- Janicke, R. U., Sprengart, M. L., Wati, M. R., and Porter, A. G.(1998). Caspase-3 is required for DNA fragmentation and morphological changes associated with apoptosis. J. Biol. Chem. 273:9357–9360.Google Scholar
- Kawasaki, T., Kunisato, A., Hazama, K., Uyeda, A., and Taguchi, T.(2001). Identification of active regions for neurite outgrowth activity of neurocrescin. Biochem. Biophys. Res. Commun. 281:761–765.Google Scholar
- Nishimune, H., Oishi, I., Koyanagi, S., and Taguchi, T.(1998). Neurite outgrowth-promoting factors in extracts of denervated chick skeletal muscle. Cell. Mol. Neurobiol. 18:391–398.Google Scholar
- Nishimune, H., Uyeda, A., Nogawa, M., Fujimori, K., and Taguchi, T.(1997). Neurocrescin: a novel neurite-outgrowth factor secreted by muscle after denervation. Neuroreport 8:3649–3654.Google Scholar
- Stenmark, H., Vitale, G., Ullrich, O., and Zerial, M.(1995). Rabaptin-5 is a direct effector of the small GTPase Rab5 in endocytic membrane fusion. Cell 83:423–432.Google Scholar
- Stennicke, H. R., and Salvesen, G. S.(1997). Biochemical characteristics of caspases-3, -6, -7, and -8. J. Biol. Chem. 272:25719–25723.Google Scholar
- Swanton, E., Bishop, N., and Woodman, P.(1999). Human rabaptin-5 is selectively cleaved by caspase-3 during apoptosis. J. Biol. Chem. 274:37583–37590.Google Scholar
- Taguchi, T., Huchet, M., Roa, M., Changeux, J. P., and Henderson, C. E.(1987). A subpopulation of embryonic telencephalic neurons survive and develop in vitro in response to factors derived from the periphery. Brain Res. 465:125–132.Google Scholar
- Tewari, M., Quan, L. T., O’Rourke, K., Desnoyers, S., Zeng, Z., Beidler, D. R., Poirier, G. G., Salvesen, G.~S., and Dixit, V. M.(1995). YamaCPP32 beta, a mammalian homolog of CED-3, is a CrmA-inhibitable protease that cleaves the death substrate poly(ADP-ribose) polymerase. Cell 81:801–809.Google Scholar
- Thornberry, N. A., Rano, T. A., Peterson, E. P., Rasper, D. M., Timkey, T., Garcia-Calvo, M., Houtzager, V. M., Nordstrom, P. A., Roy, S., Vaillancourt, J. P., Chapman, K. T., and Nicholson, D. W.(1997). A combinatorial approach defines specificities of members of the caspase family and granzyme B. Functional relationships established for key mediators of apoptosis. J. Biol. Chem. 272:17907–17911.Google Scholar
- Uyeda, A., Fukui, I., Fujimori, K., Kiyosue, K., Nishimune, H., Kasai, M., and Taguchi, T.(2000a). MDP77: A novel neurite-outgrowth-promoting protein predominantly expressed in chick muscles. Biochem. Biophy. Res. Commun. 269:564–569.Google Scholar
- Uyeda, A., Simo Wydisdtuti, M., Nishimune, H., Kiyosue, K., Kasai, M., and Taguchi, T.(2000b). Chick muscle-derived protein 62: A novel neurite outgrowth promoting protein. Neurosci. Lett. 284:61–64.Google Scholar
- Welihinda, A. A., Tirasophon, W., and Kaufman, R. J.(1999). The cellular response to protein misfolding in the endoplasmic reticulum. Gene. Expr. 7:293–300.Google Scholar