ZFIN ID: ZDB-PUB-080306-1
Zebrafish anti-apoptotic protein zfBcl-x(L) can block betanodavirus protein alpha-induced mitochondria-mediated secondary necrosis cell death
Wu, H.C., Chiu, C.S., Wu, J.L., Gong, H.Y., Chen, M.C., Lu, M.W., and Hong, J.R.
Date: 2008
Source: Fish & shellfish immunology   24(4): 436-449 (Journal)
Registered Authors: Gong, Hong-Yi, Hong, Jiann-Ruey, Wu, Jen-Leih
Keywords: Nervous necrosis virus, Protein α, Mitochondria, zfBcl-xL, Apoptosis, Secondary necrosis cell death
MeSH Terms:
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Capsid Proteins/genetics
  • Capsid Proteins/metabolism
  • Caspase 3/metabolism
  • Caspase 8/metabolism
  • Cell Line
  • Cloning, Molecular
  • Cytochromes c/metabolism
  • Enzyme Activation
  • Gene Expression Regulation
  • Membrane Potential, Mitochondrial
  • Mitochondria/metabolism*
  • Molecular Sequence Data
  • Necrosis/pathology
  • Necrosis/virology
  • Nodaviridae/physiology*
  • Viral Proteins/metabolism
  • Zebrafish/metabolism*
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism*
  • bcl-X Protein/genetics
  • bcl-X Protein/metabolism*
PubMed: 18276161 Full text @ Fish Shellfish Immunol.
ABSTRACT
Betanodavirus protein alpha 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 24h post-transfection (p.t.) by Annexin V-FLUOS staining. TUNEL assay revealed apoptotic cells at 24-72h p.t, after which post-apoptotic necrotic cells were identified by acridine orange/ethidium bromide dual dye staining from 48 to 72h p.t. Protein alpha induced progressive loss of mitochondrial membrane potential (MMP) which was detected in RNA2-transfected GF-1 cells at 24, 48, and 72h p.t., which correlated with cytochrome c release, especially at 72h p.t. To assess the effect of zfBcl-x(L) on cell death, RNA2-transfected cells were co-transfected with zfBcl-x(L). Co-transfection of GF-1 cells prevented loss of MMP at 24h and 48h p.t. and blocked initiator caspase-8 and effector caspase-3 activation at 48h p.t. We conclude that RGNNV protein alpha induces apoptosis followed by secondary necrotic cell death through a mitochondria-mediated death pathway and activation of caspases-8 and -3.
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