PUBLICATION
Zebrafish anti-apoptotic protein zfBcl-x(L) can block betanodavirus protein alpha-induced mitochondria-mediated secondary necrosis cell death
- Authors
- Wu, H.C., Chiu, C.S., Wu, J.L., Gong, H.Y., Chen, M.C., Lu, M.W., and Hong, J.R.
- ID
- ZDB-PUB-080306-1
- 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.
Citation
Wu, H.C., Chiu, C.S., Wu, J.L., Gong, H.Y., Chen, M.C., Lu, M.W., and Hong, J.R. (2008) Zebrafish anti-apoptotic protein zfBcl-x(L) can block betanodavirus protein alpha-induced mitochondria-mediated secondary necrosis cell death. Fish & shellfish immunology. 24(4):436-449.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping