Induction of apoptotic death in cells via Bad gene expression by infectious pancreatic necrosis virus infection.
A Bcl-2 related family member, Bad, promotes cell death, and its function is regulated by phosphorylation. In this study, we show how the IPNV elicits the induction of Bad gene expression and promotes host apoptotic death. Anti-IPNV-E1S polyclonal and anti-VP3 monoclonal antibodies are used to neutralize the virus that blocks the prime death signal via the virus receptor. In the viability assay, each antibody could also enhance cell viability during IPNV infection. We tested tyrosine kinase inhibitors on IPNV-infected cells in order to assess their effect on blocking the death signal. With 100 microg/ml genistein treatment, Bad-like gene expression was blocked, either by rescuing the IPNV-infected CHSE-214 cells or by blocking internucleosomal DNA cleavage; but the tyrphostin group did not block Bad expression. For CHSE-214 cells, treatment with the protein synthesis-inhibitor, cycloheximide (1microg/ml), blocked new protein synthesis via activated tyrosine kinase during IPNV infection. We found that Bad protein expression could be blocked, and apoptotic death prevented. Together, these results demonstrate that the IPNV exerts up-regulation of a pro-apoptotic death gene (Bad), the expression of which serves to trigger apoptotic cell death. Our data also suggests that the IPNV induces apoptotic death via a viral receptor which triggers death effector Bad gene expression, possibly through a tyrosine kinase-dependent pathway.
Hong
J-R
JR
Laboratory of Marine Molecular Biology and Biotechnology, Institute of Zoology, Academia Sinica, Nankang, Taipei 115, Taiwan.
Wu
J-L
JL
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D017209:Apoptosis; D015153:Blotting, Western; D002213:Capsid / Q000037:antagonists & inhibitors / Q000276:immunology; D036022:Capsid Proteins; D002352:Carrier Proteins / Q000096:biosynthesis*; D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019833:Genistein / Q000494:pharmacology; D018061:Infectious pancreatic necrosis virus / Q000276:immunology / Q000472:pathogenicity*; D007700:Kinetics; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D012474:Salmon / Q000821:virology; D014771:Virion / Q000276:immunology; D051025:bcl-Associated Death Protein
D000906:Antibodies; D036022:Capsid Proteins; D002352:Carrier Proteins; D004791:Enzyme Inhibitors; D011500:Protein Synthesis Inhibitors; D051025:bcl-Associated Death Protein; D003513:Cycloheximide; D019833:Genistein; D011505:Protein-Tyrosine Kinases