title: Beta-nodavirus B2 protein induces hydrogen peroxide production, leading to Drp1-recruited mitochondrial fragmentation and cell death via mitochondrial targeting.
abstract: Because the role of the viral B2 protein in the pathogenesis of nervous necrosis virus infection remains unknown, the aim of the present study was to determine the effects of B2 protein on hydrogen peroxide (H2O2)-mediated cell death via mitochondrial targeting. Using a B2 deletion mutant, the B2 mitochondrial targeting signal sequence ((41)RTFVISAHAA(50)) correlated with mitochondrial free radical production and cell death in fish cells, embryonic zebrafish, and human cancer cells. After treatment of grouper fin cells (GF-1) overexpressing B2 protein with the anti-oxidant drug, N-acetylcysteine (NAC), and overexpression of the antioxidant enzymes, zfCu/Zn superoxide dismutase (SOD) and zfCatalase, decreased H2O2 production and cell death were observed. To investigate the correlation between B2 cytotoxicity and H2O2 production in vivo, B2 was injected into zebrafish embryos. Cell damage, as assessed by the acridine orange assay, gradually increased over 24 h post-fertilization, and was accompanied by marked increases in H2O2 production and embryonic death. Increased oxidative stress, as evidenced by the up-regulation of Mn SOD, catalase, and Nrf2, was also observed during this period. Finally, B2-induced dynamin-related protein 1 (Drp1)-mediated mitochondrial fragmentation and cell death could be reversed by NAC and inhibitors of Drp1 and Mdivi in GF-1 cells. Taken together, betanodavirus B2 induces H2O2 production via targeting the mitochondria, where it inhibits complex II function. H2O2 activates Drp1, resulting in its association with the mitochondria, mitochondrial fission and cell death in vitro and in vivo.
authors.affiliation: Laboratory of Molecular Virology and Biotechnology, Institute of Biotechnology, National Cheng Kung University, Tainan, 701, Taiwan, ROC.
authors.forename: Yu C
authors.initials: YC
authors.lastname: Su
authors.forename: Hsuan W
authors.initials: HW
authors.lastname: Chiu
authors.forename: Jo C
authors.initials: JC
authors.lastname: Hung
authors.forename: Jiann R
authors.initials: JR
authors.lastname: Hong
mesh_terms: D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D034281:Dynamins; D005393:Fish Diseases / Q000378:metabolism* / Q000503:physiopathology / Q000821:virology; D029941:Fish Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism*; D030381:Nodaviridae / Q000235:genetics / Q000378:metabolism*; D018384:Oxidative Stress; D012327:RNA Virus Infections / Q000378:metabolism / Q000503:physiopathology / Q000662:veterinary* / Q000821:virology; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism*; D015027:Zebrafish
chemical_list: D029941:Fish Proteins; D017361:Viral Nonstructural Proteins; D006861:Hydrogen Peroxide; D034281:Dynamins