Abstract
Granzyme B is the prototypic member of a family of serine proteases localized to the cytolytic granules of cytotoxic lymphocytes. Together with another granule protein, perforin, granzyme B is capable of inducing all aspects of apoptotic death in target cells. A number of granzyme B substrates have been identified and it has been demonstrated that granzyme B is responsible, directly or indirectly, for the morphological nuclear changes observed in target cell apoptosis, including DNA fragmentation. In an earlier study, we showed that granzyme B binds to a nuclear protein in a manner dependent on its enzymatic activity. Here, we demonstrate that granzyme B is translocated rapidly to the nucleus in cells that have been induced to undergo apoptosis by a granzyme-dependent process, and that translocation is dependent on caspase activity. Appearance of granzyme B in the nucleus of target cells precedes the detection of DNA fragmentation. Although not directly responsible for DNA fragmentation, these data suggest a nuclear role for granzyme B in target cell apoptosis. c-Abl nuclear functions.
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Abbreviations
- Ad2:
-
Adenovirus type 2
- CAD:
-
caspase activated deoxyribonuclease
- CTL:
-
cytotoxic T lymphocyte
- DAPI:
-
4,6-diamidino-2-phenylindole dihydrochloride
- DCI:
-
3,4-dichloroisocoumarin
- DiOC6(3):
-
3,5′-dihexyloxacarbocyanine iodide
- DFF:
-
DNA fragmentation factor
- FITC:
-
fluorescein isothiocyanate
- ICAD:
-
inhibitor of caspase activated deoxyribonuclease
- PARP:
-
poly (ADP-ribose) polymerase
- TUNEL:
-
terminal deoxynucleotidyl transferase-mediated dUTP-FITC nick end labeling
- ΔΨm:
-
mitochondrial inner membrane potential
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Acknowledgements
MJ Pinkoski is the recipient of a Post-Doctoral Fellowship from the Medical Research Council (MRC) of Canada. JA Heilbein holds an MRC Studentship, M Barry is the recipient of a Post-Doctoral Fellowship from the Alberta Heritage Medical Foundation for Medical Research (AHFMR), and RC Bleackley is a Medical Scientist of the AHFMR, an MRC Distinguished Scientist and an International Scholar of the Howard Hughes Medical Institute. We thank Don Nicholson for the anti-caspase-3 antibody, Chris Froelich for purified granzyme B, and Prem Seth for the replication-deficient adenovirus.
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Pinkoski, M., Heibein, J., Barry, M. et al. Nuclear translocation of granzyme B in target cell apoptosis. Cell Death Differ 7, 17–24 (2000). https://doi.org/10.1038/sj.cdd.4400604
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DOI: https://doi.org/10.1038/sj.cdd.4400604
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