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Caspase-3 activates endo-exonuclease: Further evidence for a role of the nuclease in apoptosis

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Abstract

Single-strand DNase and poly rAase, activities characteristic of endo-exonuclease, were co-activated in nuclear fractions of HL-60 cells by caspase-3. Activation was accompanied by cleavages of large soluble polypeptides (130–185 kDa) and a 65 kDa inactive chromatin-associated polypeptide related to the endo-exonuclease of Neurospora crassa as detected on immunoblots. The major products seen in vitro were a 77 kDa soluble polypeptide and an active chromatin-associated 34 kDa polypeptide. When HL-60 cells were induced to undergo apoptosis by treating with 50 μM etoposide (VP-16) for 4 hours, 77 kDa and 40 kDa polypeptides accumulated in nuclear fractions. Chromatin DNA fragmentation activity was also activated in cytosol and nuclear extract either by pre-treating the cells in vivo with VP-16 or by treating the cytosol in vitro with caspase-3 or dATP and cytochrome c. Endo-exonuclease activated by caspase-3 in cytosol-derived fractions augmented chromatin DNA fragmentation activity in vitro. Endo-exonuclease is proposed to act in vivo in conjunction with the caspase-activated DNase (CAD) to degrade chromatin DNA during apoptosis of HL-60 cells.

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Authors and Affiliations

  1. Department of Immunology and Rheumatology, Sydney Children's Hospital, High St., Randwick, N. S. W., 2031, Australia

    Xue Wei Meng, J. M. Feller & J. B. Ziegler

  2. Department of Immunology and Rheumatology, Sydney Children's Hospital, High St., Randwick, N. S. W., 2031, Australia

    M. J. Fraser

Authors
  1. Xue Wei Meng
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  2. M. J. Fraser
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  3. J. M. Feller
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  4. J. B. Ziegler
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Meng, X.W., Fraser, M.J., Feller, J.M. et al. Caspase-3 activates endo-exonuclease: Further evidence for a role of the nuclease in apoptosis. Apoptosis 5, 243–254 (2000). https://doi.org/10.1023/A:1009604529237

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