Abstract
Granzyme F (GzmF) belongs to a unique group of granzymes in mice. Murine GzmF is highly expressed in NK3.1 cells and in lymphokine-activated killer (LAK) cells. However, the manner in which GzmF works in granule-mediated cytolysis is unknown. In this study, we first demonstrated that GzmF causes a novel cell death pathway. The death is characterized by an externalization of phosphatidylserine, by nuclear condensation, mitochondrial damage, cytochrome c (cyt c) release, caspase inactivation and single-stranded DNA nicking. GzmF-induced chromatin was incompletely condensed and segmented at the nuclear periphery. Cellular organelles were damaged and the cytoplasm showed an extensive vacuolization that is reminiscent of necroptosis. GzmF can cause rapid mitochondrial swelling, depolarization and reactive oxygen species accumulation. GzmF-induced death does not involve caspase activation, Bid cleavage or activation of DNA nickase NM23H1. GzmF-silenced LAK cells showed reduced cytotoxicity against caspase-inhibited target tumor cells. Moreover, cyt c release is independent of Bid or Bax/Bak. We further showed that GzmF impairs mitochondrial electron transport to abolish ATP generation. ATP decline may contribute to a failure of apoptosome formation, leading to caspase inactivation.
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Abbreviations
- Gzm:
-
granzyme
- S-AGzmF:
-
Ser195AlaGzmF
- Ad:
-
replication-deficient adenovirus type V
- PFP:
-
perforin
- SLO:
-
streptolysin O
- MEF:
-
mouse embryonic fibroblast
- STP:
-
staurosporine
- cyt c:
-
cytochrome c
- LAK:
-
lymphokine-activated killer
- shRNA:
-
small hairpin RNA
- AV:
-
annexin V
- PI:
-
propidium iodide
- ROS:
-
reactive oxygen species
- ICAD:
-
inhibitor of caspase-activated DNase
- Apaf-1:
-
apoptotic protease-activating factor 1
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Acknowledgements
We thank Dr. Q Hou and H Zhang, and D Hu and H Tang for their helpful advice; and CC Liu, L Sun and Y Teng for their technical support. Bid KO MEF is a gift from Dr. A Strasser (The Walter and Eliza Hall Institute of Medical Research, Australia); Bax/Bak DKO MEF is generously provided by Dr. Q Chen (Institute of Zoology, CAS). This work was supported from the National Natural Science Foundation of China (30525005, 30830030, 30772496, 30700126), 863 program (2006AA02Z4C9), 973 programs (2006CB504303, 2006CB910901), the Innovative program (KSCX2-YW-R-42) and the Hundred Talents Program of CAS.
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Shi, L., Wu, L., Wang, S. et al. Granzyme F induces a novel death pathway characterized by Bid-independent cytochrome c release without caspase activation. Cell Death Differ 16, 1694–1706 (2009). https://doi.org/10.1038/cdd.2009.101
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DOI: https://doi.org/10.1038/cdd.2009.101
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