Abstract
When added with cytoch rome c,Xenopus laevis egg extract XS-150 can induce exogenous nuclei undergoing apoptosis. Apoptosis specific DNase XAD was activated during this process, and cut chromatin between nucleosome, leading to DNA Ladder in electrophoresis. Our results showed that an inhibitor of XAD, IXAD, exists abundantly in normal egg extract, its molecular weight is about 40 ku. Normally, IXAD exists either in the form of dimmer or in complex with XAD. It was degraded during apoptosis, releasing active XAD. The results of Western assay and cross-inhibition showed that IXAD was likely homologous to DFF45 in structure and function. At the same time, these results also indicated that the pathway in apoptosis was conserved in evolution.
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Alnemri, E. S., Livingston, D. J., Nicholson, D. W. et al., Human ICE/CED3 protease nomenclature, Cell, 1996, 87: 171.
Thornberry, N. A., Yuri Lazebnik, Y., Caspases: Enemies within, Science, 1998, 281: 1312.
Enari, M., Sakahira, H., Yokoyama, H. et al., A caspase-activated DNase that degrades DNA during apoptosis and its inhibitor ICAD, Nature, 1998, 391: 43.
Liu, X. S., Zou, H., Slaughter, C. et al., DFF, a heterodimeric protein that functions downstream of caspase-3 to trigger DNA fragmentation during apoptosis, Cell, 1997, 89: 175.
Wolf, B. B., Schuler, M., Echeverri, F. et al., Caspase-3 is the primary activator of apoptotic DNA fragmentation via DNA fragmentation factor-45/inhibitor of caspase-activated DNase inactivation, JBC, 1999, 274: 30651.
Kluck, R. M., Martin, S. J., Hoffman, B. M. et al., Cytochrome c activation of CPP32-like proteolysis plays a critical role in a Xenopus cell-free apoptosis system, EMBO J, 1997, 16: 4639.
Zhu, S., Lu, Z. G., Sun, Y. L. et al., Specific degradation of keratin in Xenopus laevis egg extracts undergoing apoptosis, Chinese Science Bulletin, 2000, 45(21): 1977.
Zhu, S., Jiang, Z. F., Zhang, B. et al., Identification of a DNase activated in Xenopus egg extracts undergoing apoptosis, Chinese Science Bulletin, 1998, 43(6): 522.
Jiang, Z. F., Zhu, S., Zhai, Z. H., Induction of mouse nuclear apoptosis in Xenopus egg extracts, Chinese Science Bulletin, 1998, 43(4): 409.
Newmeyer, D. D., Farschon, D. M., Reed, J. C., Cell-free apoptosis in Xenopus egg extract: inhibition by Bcl-2 and requirement for an organelle fraction enriched in mitochondria, Cell, 1994, 79: 353.
Kayalar, C., Ord, T., Testa, M. P. et al., Cleavage of actin by interleukin 1β-converting enzyme to reverse DNase I inhibition, 1996, PNAS, 93: 2234.
Zou, H., Henzel, W. J., Liu, X. S. et al., Apaf-1, a human protein homologous to C. elegans CED-4, participates in cytochrome c-dependent activation of caspase-3, Cell, 1997, 90: 405.
McCall, K., Steller, H., Requirement for DCP-1 caspase during Drosophila oogenesis, Science, 1998, 279: 230.
Walker, P. R., Leblanc, J., Carson, C. et al., Neither caspase-3 nor DNA fragmentation factor is required for high molecular weight DNA degradation in apoptosis, Ann. N. Y. Acad. Sci., 1999, 887: 48.
Clarke, P. G. H., Developmental cell death: morphological diversity and multiple mechanisms, Anat. Embryol., 1990, 181: 195.
Nakamura, N., Wada, Y., Properties of DNA fragmentation activity generated by ATP depletion, Cell Death Differ., 2000, 7(5): 477.
Liu, Q. Y., Ribecco, M., Pandey, S. et al., Apoptosis-related functional features of the DNaseI-like family of nucleases, Ann. N. Y. Acad. Sci., 1999, 887: 60.
Zhang, J., Lee, H., Lou, D. W. et al., Lack of obvious 50 kilobase pair DNA fragments in DNA fragmentation factor 45-deficient thymocytes upon activation of apoptosis, Biochem. Biophys. Res. Commun., 2000, 274(1): 225.
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Lu, Z., Yang, W., Cao, Q. et al. Identification of inhibitor of apoptosis specific DNase in Xenopus egg extract. Chin.Sci.Bull. 46, 1449–1453 (2001). https://doi.org/10.1007/BF03187029
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DOI: https://doi.org/10.1007/BF03187029