title: Endogenous grouper and barramundi Mx proteins facilitated the clearance of betanodavirus RNA-dependent RNA polymerase.
abstract: This study confirmed that the infection of nervous necrosis virus (NNV), belonging to the betanodavirus, can induce the expression of endogenous Mx in grouper fin-3 (GF-3), grouper brain (cGB), and barramundi brain (cBB) cells, but not in grouper fin-1 (GF-1) cells. In a co-sedimentation assay, RdRp appeared in the mitochondrial pellet of GF-1 cells without endogenous Mx expression. However, in GF-3, cGB, and cBB cells, RdRp was detected in the nuclear pellet accompanied by endogenous Mx. By immunostaining, RdRp was found to colocalize with not only endogenous Mx but also lysosomes and monodansylcadaverine (MDC)-labeled autophagic vacuoles. In GF-1 cells, the RdRp level continuously increased during 24-72 h post infection (hpi). When endogenous Mx expressed during 24-72 hpi in virus-infected GF-3, cGB, and cBB cells, the RdRp level peaked at 24 hpi but decreased at 48-72 hpi. The degradation of RdRp could be suppressed by treatment with 3-methyladenine (3MA), NH4Cl, and Mx-specific siRNA respectively. After poly I:C transfection, the endogenous Mx level peaked at 3 days post transfection (dpt) and then spontaneously decreased at 5-7 dpt. The poly I:C-indued Mx also colocalized with MDC-labeled autophagic vacuoles at 3 dpt, and its degradation could be inhibited by 3MA or NH4Cl treatments. Therefore, the anti-NNV mechanism of endogenous grouper and barramundi Mx is suggested to sequester RdRp for degradation through autophagy and lysosomes.
authors.affiliation: Department of Life Science, National Taiwan University, Taipei, Taiwan; Institute of Molecular Medicine, National Taiwan University, Taipei, Taiwan.
authors.forename: Y-C
authors.initials: YC
authors.lastname: Wu
authors.affiliation: Department of Life Science, National Taiwan University, Taipei, Taiwan.
authors.forename: P-Y
authors.initials: PY
authors.lastname: Tsai
authors.affiliation: Department of Life Science, National Taiwan University, Taipei, Taiwan.
authors.forename: J-C
authors.initials: JC
authors.lastname: Chan
authors.affiliation: Department of Life Science, National Taiwan University, Taipei, Taiwan. Electronic address: shauchi@ntu.edu.tw.
authors.forename: S-C
authors.initials: SC
authors.lastname: Chi
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000595:Amino Acid Sequence; D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D001343:Autophagy / Q000276:immunology; D001921:Brain / Q000166:cytology / Q000378:metabolism; D002460:Cell Line; D005393:Fish Diseases / Q000276:immunology* / Q000821:virology; D008928:Mitochondria / Q000276:immunology* / Q000378:metabolism; D064547:Myxovirus Resistance Proteins / Q000235:genetics / Q000378:metabolism*; D030381:Nodaviridae / Q000276:immunology*; D010471:Perches / Q000276:immunology* / Q000821:virology; D011070:Poly I-C / Q000378:metabolism; D034622:RNA Interference; D012327:RNA Virus Infections / Q000276:immunology / Q000821:virology; D034741:RNA, Small Interfering / Q000235:genetics; D012324:RNA-Dependent RNA Polymerase; D014779:Virus Replication / Q000276:immunology
chemical_list: D064547:Myxovirus Resistance Proteins; D034741:RNA, Small Interfering; D000643:Ammonium Chloride; C025946:3-methyladenine; D012324:RNA-Dependent RNA Polymerase; D000225:Adenine; D011070:Poly I-C
keywords: Autophagy; Barramundi; Betanodavirus; Grouper; Lysosomes; Mx