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Pavithiran, Amirthalingam,Bathige, S.D.N.K.,Kugapreethan, Roopasingam,Priyathilaka, Thanthrige Thiunuwan,Yang, Hyerim,Kim, Myoung-Jin,Lee, Jehee Elsevier 2018 Fish & shellfish immunology Vol.74 No.-
<P><B>Abstract</B></P> <P>Akirins, members of the NF-κB signaling pathway, are highly conserved nuclear proteins, which regulate gene expression in many physiological processes, including immunity, myogenesis, carcinogenesis, and embryogenesis. The akirin family in teleost fish consists of two to three genes. In the present study, three akirin genes from <I>Hippocampus abdominalis</I> were identified from a transcriptome database and designated as <I>HaAkirin1</I>, <I>HaAkirin2(1)</I>, and <I>HaAkirin2(2)</I>. The nuclear localization of <I>HaAkirin1</I> and <I>HaAkirin2(1)</I> was confirmed by subcellular localization analysis. In contrast, diffused localization of <I>HaAkirin2(2)</I> was identified in the nucleus and cytoplasm that confirmed the aberrant nature of the nuclear localization signal. Phylogenetic analysis revealed a closer relationship of HaAkirins with other known teleost akirins. All three <I>HaAkirin</I> transcripts were ubiquitously expressed in all examined tissues with higher expression in ovary tissue. Immune challenge with LPS, poly I:C, and <I>Streptococcus iniae</I> exhibited a significant increase in the expression of all three <I>HaAkirins</I> in kidney and liver tissues. NF-κB luciferase assays revealed that relative luciferase activity was significantly higher for all three <I>HaAkirin</I> genes than mock controls. These results suggest that <I>HaAkirin</I> genes might play a role in regulating NF-κB dependent immune gene expression and their expression could be induced by bacterial and viral pathogen recognition molecular patterns.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Three akirin genes (<I>Haakirins</I>) were identified from <I>Hippocampus abdominalis</I>. </LI> <LI> The loss of second nuclear localization signal was identified in <I>Haakirin2(2).</I> </LI> <LI> All three <I>Haakirins</I> were prominently expressed in gonadal tissues. </LI> <LI> All three <I>Haakirins</I> were upregulated after challenging with LPS, poly I:C, and <I>S. iniae.</I> </LI> <LI> The expression of NF-κB reporter gene was induced by all three <I>Haakirins.</I> </LI> </UL> </P>
Tharuka, M.D. Neranjan,Priyathilaka, Thanthrige Thiunuwan,Yang, Hyerim,Pavithiran, Amirthalingam,Lee, Jehee ACADEMIC PRESS LTD 2019 FISH AND SHELLFISH IMMUNOLOGY Vol.86 No.-
<P><B>Abstract</B></P> <P>Viperin is recognized as an antiviral protein that is stimulated by interferon, viral exposures, and other pathogenic molecules in vertebrate. In this study, a viperin homolog in the Big-belly seahorse (<I>Hippocampus abdominalis</I>; HaVip) was functionally characterized to determine its subcellular localization, expression pattern, and antiviral activity <I>in vitro</I>. The <I>HaVip</I> coding sequence encodes a 348 amino acid polypeptide with predicted molecular weight of 38.48 kDa. Sequence analysis revealed that HaVip comprises three main domains: the N-terminal amphipathic α-helix, a radical S-adenosyl-<SMALL>L</SMALL>-methionine (SAM) domain, and a conserved C-terminal domain. Transfected GFP-tagged HaVip protein was found to localize to the endoplasmic reticulum (ER). Overexpressed-HaVip in FHM cells was found to significantly reduce viral capsid gene expression in VHSV infection <I>in vitro</I>. Under normal physiological conditions, <I>HaVip</I> expression was ubiquitously detected in all 14 examined tissues of the seahorse, with the highest expression observed in the heart, followed by skin and blood. <I>In vivo</I> studies showed that <I>HaV</I>ip was rapidly and predominantly upregulated in blood, kidney, and intestinal tissue upon poly (I:C) stimulus. LPS and <I>Streptococus iniae</I> challenges caused a significant increase in expression of <I>HaVip</I> in all the analyzed tissues. The obtained results suggest that HaVip is involved in the immune system of the seahorse, triggering antiviral and antibacterial responses, upon viral and bacterial pathogenic infections.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Identified and characterized viperin from big-belly seahorse. </LI> <LI> The HaVip gene was ubiquitously expressed in unchallenged-fish tissues. </LI> <LI> Modulated HaVip transcription pattern revealed its contribution in the immune response. </LI> <LI> Overexpression of HaVip in FHM cells showed the antiviral activity. </LI> </UL> </P>