DNA STRAND BREAKS OCCURRING DURING APOPTOSIS - THEIR EARLY INSITU DETECTION BY THE TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE AND NICK TRANSLATION ASSAYS AND PREVENTION BY SERINE PROTEASE INHIBITORS

  • Authors:
    • W GORCZYCA
    • S BRUNO
    • RJ DARZYNKIEWICZ
    • JP GONG
    • Z DARZYNKIEWICZ
  • View Affiliations

  • Published online on: November 1, 1992     https://doi.org/10.3892/ijo.1.6.639
  • Pages: 639-648
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Abstract

The appearance of DNA strand breaks during apoptosis was detected in individual cells, in relation to the cell cycle phase, by a novel assay based on labeling 3'-OH termini with biotinylated dUTP using exogenous terminal transferase or DNA polymerase. Apoptosis was induced in HL-60 cells by the DNA topoisomerase I and II inhibitors, and in rat thymocytes by prednisolone. Formation of strand breaks was prevented by the serine protease irreversible inhibitors diisopropyl fluorophosphate (DFP), L-1-tosylamido-2-phenylethyl chloromethyl ketone (TPCK), N-p-tosyl-L-lysine chloromethyl ketone (TLCK) and by the substrates N-alpha-tosyl-L-arginine methyl ester (TAME) and N-benzoyl-L-tyrosine ethyl ester (BTEE). The data indicate that initiation of DNA degradation during apoptosis is preceded by a proteolytic step and suggest that apoptosis starts with activation (e.g. by DNA lesions) of a serine protease which hydrolyses protein(s) associated with the internucleosomal linker DNA sections, thus increasing accessibility of linker DNA to the apoptosis-associated endonuclease.

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November 1992
Volume 1 Issue 6

Print ISSN: 1019-6439
Online ISSN:1791-2423

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Spandidos Publications style
GORCZYCA W, BRUNO S, DARZYNKIEWICZ R, GONG J and DARZYNKIEWICZ Z: DNA STRAND BREAKS OCCURRING DURING APOPTOSIS - THEIR EARLY INSITU DETECTION BY THE TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE AND NICK TRANSLATION ASSAYS AND PREVENTION BY SERINE PROTEASE INHIBITORS. Int J Oncol 1: 639-648, 1992.
APA
GORCZYCA, W., BRUNO, S., DARZYNKIEWICZ, R., GONG, J., & DARZYNKIEWICZ, Z. (1992). DNA STRAND BREAKS OCCURRING DURING APOPTOSIS - THEIR EARLY INSITU DETECTION BY THE TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE AND NICK TRANSLATION ASSAYS AND PREVENTION BY SERINE PROTEASE INHIBITORS. International Journal of Oncology, 1, 639-648. https://doi.org/10.3892/ijo.1.6.639
MLA
GORCZYCA, W., BRUNO, S., DARZYNKIEWICZ, R., GONG, J., DARZYNKIEWICZ, Z."DNA STRAND BREAKS OCCURRING DURING APOPTOSIS - THEIR EARLY INSITU DETECTION BY THE TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE AND NICK TRANSLATION ASSAYS AND PREVENTION BY SERINE PROTEASE INHIBITORS". International Journal of Oncology 1.6 (1992): 639-648.
Chicago
GORCZYCA, W., BRUNO, S., DARZYNKIEWICZ, R., GONG, J., DARZYNKIEWICZ, Z."DNA STRAND BREAKS OCCURRING DURING APOPTOSIS - THEIR EARLY INSITU DETECTION BY THE TERMINAL DEOXYNUCLEOTIDYL TRANSFERASE AND NICK TRANSLATION ASSAYS AND PREVENTION BY SERINE PROTEASE INHIBITORS". International Journal of Oncology 1, no. 6 (1992): 639-648. https://doi.org/10.3892/ijo.1.6.639