Molecular cloning and expression analysis of three Mx isoforms of rock bream, Oplegnathus fasciatus

Zenke, Kosuke; Kim, Ki Hong

doi:10.1016/j.fsi.2008.09.012

Cited by 22 publications

(11 citation statements)

References 28 publications

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“…The genomic organization of rsbMx1 and rsbMx2 comprised 12 exons and 11 introns. This is agreement with other fishes as rock bream Mx1, Mx2, Mx3 and pufferfish Mx genes contain 11, 11, 12 and 12 exons, respectively (Yap et al 2003;Zenke & Kim 2009). To evaluate and compare the innate immune response in splenocytes, red sea bream and rock bream were infected with IVS-1 and analysed the immune gene expression at 1, 3, and 7 days post-injection.…”

Section: Discussionsupporting

confidence: 90%

“…Although the isomers of Mx were identified in red sea bream and rock bream, we analysed the conserved region of Mx to detect Mx1 and Mx2 of red sea bream and Mx1, Mx2 and Mx3 of rock bream simultaneously. This is because Zenke & Kim (2009) reported that the gene expression patterns of Mx1 and Mx2 following poly I:C stimulation were similar.…”

Section: Expression Of Innate Immune Genes In Spleen In Vivomentioning

confidence: 81%

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Cloning and expression analysis of innate immune genes from red sea bream to assess different susceptibility to megalocytivirus infection

Jin

Kim

Hong

et al. 2016

Journal of Fish Diseases

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As suggested by the Office International des Epizooties (OIE), fishes belonging to the genus Oplegnathus are more sensitive to megalocytivirus infection than other fish species including red sea bream (Pagrus major). To assess the roles of the innate immune response to these different susceptibilities, we cloned the genes encoding inflammatory factors including IL-8 and COX-2, and the antiviral factor like Mx from red sea bream for the first time and performed phylogenetic and structural analysis. Analysed expression levels of IL-1β, IL-8 and COX-2 and the antiviral factor like Mx genes performed with in vivo challenge experiment showed no difference in inflammatory gene expression or respiratory burst activity between red sea bream and rock bream (Oplegnathus fasciatus). However, the Mx gene expression levels in red sea bream were markedly higher than those in rock bream, suggesting the importance of type I interferon (IFN)-induced proteins, particularly Mx, during megalocytivirus infection, rather than inflammation-related genes. The in vitro challenge experiments using embryonic primary cultures derived from both fish species showed no difference in cytopathic effects (CPE), viral replication profiles, and inflammatory and Mx gene expression pattern between the two fish species.

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Section: Discussionsupporting

confidence: 90%

Section: Expression Of Innate Immune Genes In Spleen In Vivomentioning

confidence: 81%

Cloning and expression analysis of innate immune genes from red sea bream to assess different susceptibility to megalocytivirus infection

Jin

Kim

Hong

et al. 2016

Journal of Fish Diseases

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“…They belong to the class of dynamin-like large GTPases known to be involved in intracellular vesicle trafficking and organelle homeostasis [11]. Generally, the N-terminal region is highly conserved and thought to have a regulatory function, whereas the C-terminal region is divergent and acts as an effector domain [25], which facilitates defense against a diverse range of viruses. The N-terminal GTPase domain binds and hydrolyses GTP [14], and the C-terminal LZ plays a role in the aggregation of dimers and trimers [26].…”

Section: Discussionmentioning

confidence: 99%

Protective Roles of Grass Carp Ctenopharyngodon idella Mx Isoforms against Grass Carp Reovirus

Peng

Yang

2012

PLoS ONE

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BackgroundMyxovirus 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.Methodology/Principal FindingsIn 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.Conclusions/SignificanceThe 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.

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“…The primers are designed based on the reported Mx, IL‐ 1β and IL‐ 8 sequences of rock bream. Each assay was performed in triplicate with β‐actin mRNA as the control as described previously (Zenke & Kim 2009). All data are given in terms of relative mRNA, expressed as means plus or minus standard errors of the means.…”

Section: Methodsmentioning

confidence: 99%

Characterization of a megalocytivirus from cultured rock bream, Oplegnathus fasciatus (Temminck & Schlege), in China

Zhang

Xiao

et al. 2011

Aquaculture Research

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Megalocytivirus belongs to the Iridoviridae family and is known to a¡ect ¢n¢sh. Megalocytivirus epizootics have been reported to occur in several cultured ¢sh species in China; however, no megalocytivirus associated with rock bream, Oplegnathus fasciatus (Temminck & Schlege), has been documented. In this study, we characterized for the ¢rst time a megalocytivirus, rock bream iridovirus (RBIV)-C1, detected in cultured rock bream in a ¢sh farm in China that had been in£icted with a high-mortality disease outbreak. Sequence analysis of three conserved genes showed that RBIV-C1 shares over 90% overall identities with a number of known megalocytiviruses. Electron microscopic examination revealed RBIV-C1 as hexagonal particles similar to those reported for megalocytiviruses. In vivo infection study indicated that, following inoculation into rock bream, RBIV-C1induced100% mortality and upregulated the expression of Mx, IL-1b and IL-8. The infected ¢sh exhibited pathological signs similar to those observed in naturally diseased ¢sh. Furthermore, studies in a turbot (Scophthalmus maximus L.) model indicated that RBIV-C1 induced acute infection in turbot that led to 100% mortality. These results indicate that RBIV-C1 is highly virulent to rock bream as well as turbot and that RBIV-C1 is closely related to a number of previously reported megalocytivirus and likely a genetic variant of the latter.

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Molecular cloning and expression analysis of three Mx isoforms of rock bream, Oplegnathus fasciatus

Cited by 22 publications

References 28 publications

Cloning and expression analysis of innate immune genes from red sea bream to assess different susceptibility to megalocytivirus infection

Cloning and expression analysis of innate immune genes from red sea bream to assess different susceptibility to megalocytivirus infection

Protective Roles of Grass Carp Ctenopharyngodon idella Mx Isoforms against Grass Carp Reovirus

Characterization of a megalocytivirus from cultured rock bream, Oplegnathus fasciatus (Temminck & Schlege), in China

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