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PloS one, 2012-12, Vol.7 (12), p.e52142-e52142
2012
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Autor(en) / Beteiligte
Titel
Protective roles of grass carp Ctenopharyngodon idella Mx isoforms against grass carp reovirus
Ist Teil von
  • PloS one, 2012-12, Vol.7 (12), p.e52142-e52142
Ort / Verlag
United States: Public Library of Science
Erscheinungsjahr
2012
Quelle
MEDLINE
Beschreibungen/Notizen
  • Myxovirus resistance (Mx) proteins are crucial effectors of the innate antiviral response against a wide range of viruses, mediated by the type I interferon (IFN-I) signaling pathway. However, the antiviral activity of Mx proteins is diverse and complicated in different species. In the current study, two novel Mx genes (CiMx1 and CiMx3) were identified in grass carp (Ctenopharyngodon idella). CiMx1 and CiMx3 proteins exhibit high sequence identity (92.1%), and low identity with CiMx2 (49.2% and 49.5%, respectively) from the GenBank database. The predicted three-dimensional (3D) structures are distinct among the three isoforms. mRNA instability motifs also display significant differences in the three genes. The spatial and temporal expression profiles of three C. idella Mx genes and the IFN-I gene were investigated by real-time fluorescence quantitative RT-PCR (qRT-PCR) following infection with grass carp reovirus (GCRV) in vivo and in vitro. The results demonstrated that all the four genes were implicated in the anti-GCRV immune response, that mRNA expression of Mx genes might be independent of IFN-I, and that CIK cells are suitable for antiviral studies. By comparing expression patterns following GCRV challenge or poly(I:C) treatment, it was observed that GCRV blocks mRNA expression of the four genes. To determine the functions of Mx genes, three CiMx cDNAs were cloned into expression vectors and utilized for transfection of CIK cells. The protection conferred by each recombinant CiMx protein against GCRV infection was evaluated. Antiviral activity against GCRV was demonstrated by reduced cytopathic effect, lower virus titer and lower levels of expressed viral transcripts. The transcription of IFN-I gene was also monitored. The results indicate all three Mx genes can suppress replication of grass carp reovirus and over-expression of Mx genes mediate feedback inhibition of the IFN-I gene.
Sprache
Englisch
Identifikatoren
ISSN: 1932-6203
eISSN: 1932-6203
DOI: 10.1371/journal.pone.0052142
Titel-ID: cdi_plos_journals_1327146415
Format
Schlagworte
Amino Acid Sequence, Amino acids, Animal sciences, Animals, Antiviral activity, Biological response modifiers, Biology, Carp, Carps, Carps - genetics, Carps - immunology, Carps - metabolism, Carps - virology, Ctenopharyngodon idella, Cytokines, DNA, Complementary - genetics, Expression vectors, Feedback inhibition, Fish, Fish Diseases - genetics, Fish Diseases - immunology, Fish Diseases - metabolism, Fish Diseases - virology, Fishes, Fluorescence, Gene expression, Genes, Genetic engineering, Gobiocypris rarus, GTP-Binding Proteins - genetics, GTP-Binding Proteins - immunology, GTP-Binding Proteins - metabolism, Health aspects, Immune response, Immune system, Infections, Influenza, Interferon, Interferon Type I - genetics, Interferon Type I - immunology, Interferon Type I - metabolism, Isoforms, Kinases, Laboratories, Lates calcarifer, Mammals, Messenger RNA, Molecular biology, Molecular Sequence Data, Myxovirus Resistance Proteins, Next-generation sequencing, Oncorhynchus mykiss, Overexpression, Pathogens, Phylogeny, Poly (I:C), Polymerase chain reaction, Protein Isoforms, Proteins, Reoviridae - immunology, Reoviridae Infections - immunology, Reoviridae Infections - metabolism, Reoviridae Infections - veterinary, Reoviridae Infections - virology, RNA Stability, Sequence Alignment - methods, Signal transduction, Signaling, Structural stability, Studies, Transcription, Transcription (Genetics), Transfection, Trout, Viral infections, Virus Replication - genetics, Virus Replication - immunology, Viruses, Zebrafish, Zoology

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