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Navaneethaiyer Umasut,SDNK Bathige,William Shanthakumar Thulasitha,Minyoung Oh,Jehee Lee 제주대학교 해양과학연구소 2020 해양과환경연구소 연구논문집 Vol.44 No.-
CXCL8 (interleukin-8, IL-8) is a CXC family chemokine that recruits specific target cells and mediates inflammation and wound healing. This study reports the identification and characterization of two cxcl8 homologs from rock bream, Oplegnathus fasciatus. Investigation of molecular signature, homology, phylogeny, and gene structure suggested that they belonged to lineages 1 (L1) and 3 (L3), and designated Ofcxcl8-L1 and Ofcxcl8-L3. While Ofcxcl8-L1 and Ofcxcl8-L3 revealed quadripartite and tripartite organization, in place of the mammalian ELR (Glu-Leu-Arg) motif, their peptides harbored EMH (Glu-Met-His) and NSH (Asn-Ser-His) motifs, respectively. Transcripts of Ofcxcl8s were constitutively detected by Quantitative Real-Time PCR (qPCR) in 11 tissues examined, however, at dierent levels. Ofcxcl8-L1 transcript robustly responded to treatments with stimulants, such as flagellin, concanavalin A, lipopolysaccharide, and poly(I:C), and pathogens, including Edwardsiella tarda, Streptococcus iniae, and rock bream iridovirus, when compared with Ofcxcl8-L3 mRNA. The dierences in the putative promoter features may partly explain the dierential transcriptional modulation of Ofcxcl8s. Purified recombinant OfCXCL8 (rOfCXCL8) proteins were used in in vitro chemotaxis and proliferation assays. Despite the lack of ELR motif, both rOfCXCL8s exhibited leukocyte chemotactic and proliferative functions, where the potency of rOfCXCL8-L1 was robust and significant compared to that of rOfCXCL8-L3. The results, taken together, are indicative of the crucial importance of Ofcxcl8s in inflammatory responses and immunoregulatory roles in rock bream immunity.
Umasuthan, Navaneethaiyer,Bathige, S.D.N.K.,Whang, Ilson,Lim, Bong-Soo,Choi, Cheol Young,Lee, Jehee Elsevier 2015 FISH AND SHELLFISH IMMUNOLOGY Vol.43 No.2
<P><B>Abstract</B></P> <P>As a pivotal signaling mediator of toll-like receptor (TLR) and interleukin (IL)-1 receptor (IL-1R) signaling cascades, the IL-1R-associated kinase 4 (IRAK4) is engaged in the activation of host immunity. This study investigates the molecular and expressional profiles of an <I>IRAK4-like</I> homolog from <I>Oplegnathus fasciatus</I> (<I>OfIRAK4</I>). The <I>OfIRAK4</I> gene (8.2?kb) was structured with eleven exons and ten introns. A putative coding sequence (1395bp) was translated to the OfIRAK protein of 464 amino acids. The deduced OfIRAK4 protein featured a bipartite domain structure composed of a death domain (DD) and a kinase domain (PKc). Teleost IRAK4 appears to be distinct and divergent from that of tetrapods in terms of its exon-intron structure and evolutionary relatedness. Analysis of the sequence upstream of translation initiation site revealed the presence of putative regulatory elements, including NF-κB-binding sites, which are possibly involved in transcriptional control of <I>OfIRAK4</I>. Quantitative real-time PCR (qPCR) was employed to assess the transcriptional expression of <I>OfIRAK4</I> in different juvenile tissues and post-injection of different immunogens and pathogens. Ubiquitous basal mRNA expression was widely detected with highest level in liver. <I>In?vivo</I> flagellin (FLA) challenge significantly intensified its mRNA levels in intestine, liver and head kidney indicating its role in FLA-induced signaling. Meanwhile, up-regulated expression was also determined in liver and head kidney of animals challenged with potent immunogens (LPS and poly I:C) and pathogens (<I>Edwardsiella tarda</I> and <I>Streptococcus iniae</I> and rock bream iridovirus (RBIV)). Taken together, these data implicate that <I>OfIRAK4</I> might be engaged in antibacterial and antiviral immunity in rock bream.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Rock bream <I>IRAK4</I> (<I>OfIRAK4</I>) had 11 exons encoding its protein with a bipartite domain structure. </LI> <LI> Teleost IRAK4s are distinct from tetrapod IRAK4s in terms of gene structure and phylogeny. </LI> <LI> Liver was the primary site of <I>OfIRAK4</I>'s transcription. </LI> <LI> <I>OfIRAK4</I>'s putative role in flagellin sensing was evidenced from its transcriptional response. </LI> <LI> Immunogenic PAMPs/pathogens induced <I>OfIRAK4</I>'s expression implied its association with immunity. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>
Umasuthan, Navaneethaiyer,Whang, Ilson,Saranya Revathy, Kasthuri,Oh, Myung-Joo,Jung, Sung-Ju,Choi, Cheol Young,Lee, Jeong-Ho,Noh, Jae Koo,Lee, Jehee Elsevier 2012 FISH AND SHELLFISH IMMUNOLOGY Vol.32 No.5
<P><B>Abstract</B></P><P>Angiotensinogen (AGT) is the precursor of the renin-angiotensin system and contributes to osmoregulation, acute-phase and immune responses. A full-length cDNA of the <I>AGT</I> (2004 bp with a 1389 bp coding region) was isolated from rock bream (Rb), <I>Oplegnathus fasciatus</I>. The encoded polypeptide of 463 amino acids had a predicted molecular mass of 51.6 kDa. <I>RbAGT</I> possessed a deduced signal peptide of 22 residues upstream of a putative angiotensin I sequence (<SUP>23</SUP>NRVYVHPFHL<SUP>32</SUP>). <I>RbAGT</I> possessed a specific domain profile and a signature motif which are characteristics of the serpin family. Sequence homology and phylogenetic analysis indicated that <I>RbAGT</I> was evolutionarily closest to AGT of <I>Rhabdosargus sarba</I>. The mRNA expression profile of <I>RbAGT</I> was determined by quantitative RT-PCR and it demonstrated a constitutive and tissue-specific expression with the highest transcript level in the liver. Significantly up-regulated <I>RbAGT</I> expression was elicited by systemic injection of a lipopolysaccharide, rock bream iridovirus (RBIV) and bacteria (<I>Edwardsiella tarda</I> and <I>Streptococcus iniae</I>), revealing its pathogen inducibility. <I>RbAGT</I> manifested a down-regulated response to systemic injury, contemporaneously with two other serpins, protease nexin-1 (<I>PN-</I>1), and heparin cofactor II (<I>HCII</I>). In addition, a synchronized expression pattern was elicited by <I>RbAGT</I> and <I>RbTNF-α</I> in response to injury, suggesting that TNF-α might be a potential modulator of <I>AGT</I> transcription.</P> <P><B>Graphical abstract</B></P><P><ce:figure id='dfig1'></ce:figure></P><P><B>Highlights</B></P><P>► Molecular characterization of angiotensinogen from rock bream (<I>RbAGT</I>). ► Tissue-specific transcriptional profile of <I>RbAGT.</I> ► Response of hepatic <I>RbAGT</I> against LPS, bacteria and iridovirus. ► Temporal expression of hematic <I>RbAGT</I> upon injury. ► Expressional relationship between <I>RbAGT</I>, <I>HCII</I>, <I>PN-1</I> and <I>TNF-α</I>.</P>