Apoptosis

, Volume 11, Issue 9, pp 1473–1487 | Cite as

Caspase-activated DNase (CAD)-independent oligonucleosomal DNA fragmentation in chronic myeloid leukaemia cells; a requirement for serine protease and Mn2+-dependent acidic endonuclease activity

  • L. B. McGrath
  • V. Onnis
  • G. Campiani
  • D. C. Williams
  • D. M. Zisterer
  • M. M. Mc Gee
Article

Abstract

We have previously reported that the pro-apoptotic pyrrolobenzoxazepine, PBOX-6, induces apoptosis in chronic myelogenous leukaemia (CML) cells which is accompanied by oligonucleosomal DNA fragmentation. In this study we show that PBOX-6-induced oligonucleosomal DNA fragmentation occurs in the absence of caspase and CAD activation in CML cells. Dissection of the signalling pathway has revealed that induction of apoptosis requires the upstream activation of a trypsin-like serine protease that promotes the phosphorylation and inactivation of anti-apoptotic Bcl-2. In addition, in this system chymotrypsin-like serine proteases are dispensable for high molecular weight DNA fragmentation, however are required for the activation of a relatively small manganese-dependent acidic endonuclease that is responsible for oligonucleosomal fragmentation of DNA. Furthermore, we demonstrate mitochondrial involvement during PBOX-6-induced apoptosis and suggest the existence of unidentified mitochondrial effectors of apoptosis.

Keywords

Caspase-independent Pyrrolobenzoxazepine Apoptosis Serine proteases Endonuclease 

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Copyright information

© Springer Science + Business Media, LLC 2006

Authors and Affiliations

  • L. B. McGrath
    • 1
  • V. Onnis
    • 2
  • G. Campiani
    • 3
  • D. C. Williams
    • 2
  • D. M. Zisterer
    • 2
  • M. M. Mc Gee
    • 1
  1. 1.UCD School of Biomolecular and Biomedical Science, Conway InstituteUniversity College DublinDublin 4Ireland
  2. 2.School of Biochemistry and ImmunologyTrinity CollegeDublin 2Ireland
  3. 3.Dipartimento Farmaco Chimico TecnologicoUniversita'delgi Studi di SienaSienaItaly

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