Zebrafish anti-apoptotic protein zfBcl-xL can block betanodavirus protein α-induced mitochondria-mediated secondary necrosis cell death

Horng Cherng Wu, Chuan Sheng Chiu, Jen Leih Wu, Hong Yi Gong, Ming Chyuan Chen, Ming Wei Lu, Jiann Ruey Hong

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31 Citations (Scopus)


Betanodavirus protein α induces cell apoptosis or secondary necrosis by a poorly understood process. In the present work, red spotted grouper nervous necrosis virus (RGNNV) RNA 2 was cloned and transfected into tissue culture cells (GF-1) which then underwent apoptosis or post-apoptotic necrosis. In the early apoptotic stage, progressive phosphatidylserine externalization was evident at 24 h post-transfection (p.t.) by Annexin V-FLUOS staining. TUNEL assay revealed apoptotic cells at 24-72 h p.t, after which post-apoptotic necrotic cells were identified by acridine orange/ethidium bromide dual dye staining from 48 to 72 h p.t. Protein α induced progressive loss of mitochondrial membrane potential (MMP) which was detected in RNA2-transfected GF-1 cells at 24, 48, and 72 h p.t., which correlated with cytochrome c release, especially at 72 h p.t. To assess the effect of zfBcl-xL on cell death, RNA2-transfected cells were co-transfected with zfBcl-xL. Co-transfection of GF-1 cells prevented loss of MMP at 24 h and 48 h p.t. and blocked initiator caspase-8 and effector caspase-3 activation at 48 h p.t. We conclude that RGNNV protein α induces apoptosis followed by secondary necrotic cell death through a mitochondria-mediated death pathway and activation of caspases-8 and -3.

Original languageEnglish
Pages (from-to)436-449
Number of pages14
JournalFish and Shellfish Immunology
Issue number4
Publication statusPublished - 2008 Apr

All Science Journal Classification (ASJC) codes

  • Environmental Chemistry
  • Aquatic Science


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