Molecular cloning and functional characterization of two duplicated two-cysteine containing type I interferon genes in rock bream Oplegnathus fasciatus

Two type I interferon (IFN) genes, designated as rbIFN1 and rbIFN2, have been cloned and characterized in rock bream. They are both comprised of 5 exons and 4 introns, and are closely linked on the rock bream chromosome in a unique head-to-head configuration. Both genes encode 183 amino acid (aa) pr...

Full description

Saved in:

Bibliographic Details
Published in:	Fish & shellfish immunology Vol. 33; no. 4; pp. 886 - 898
Main Authors:	Wan, Qiang, Wicramaarachchi, W.D. Niroshana, Whang, Ilson, Lim, Bong-Soo, Oh, Myung-Joo, Jung, Sung-Ju, Kim, Hyun Chul, Yeo, Sang-Yeob, Lee, Jehee
Format:	Journal Article
Language:	English
Published:	England Elsevier Ltd 01-10-2012
Subjects:	Amino Acid Sequence Animals Base Sequence Blood Cells - immunology Blood Cells - metabolism Chromosomes, Artificial, Bacterial - genetics Cloning, Molecular Cysteine - genetics Cysteine - immunology Danio rerio Dicentrarchus labrax DNA Virus Infections - immunology DNA Virus Infections - veterinary Edwardsiella tarda Edwardsiella tarda - physiology Enterobacteriaceae Infections - immunology Enterobacteriaceae Infections - veterinary Escherichia coli Escherichia coli - genetics Fish Proteins - genetics Fish Proteins - immunology Freshwater Gene Duplication Genomic Library GTP-Binding Proteins - chemistry GTP-Binding Proteins - genetics Head Kidney - immunology Head Kidney - metabolism Immunity, Innate Interferon Type I - genetics Interferon Type I - immunology Iridoviridae - physiology Iridovirus Lipopolysaccharides - immunology Lipopolysaccharides - pharmacology Molecular Sequence Data Myxovirus Resistance Proteins Oplegnathus fasciatus Organ Specificity Perciformes - genetics Perciformes - immunology Phylogeny Poly I-C - immunology Real-Time Polymerase Chain Reaction - veterinary Reverse Transcriptase Polymerase Chain Reaction - veterinary Rock bream Sequence Alignment - veterinary Sequence Homology, Amino Acid Streptococcal Infections - immunology Streptococcal Infections - veterinary Streptococcus - physiology Streptococcus iniae Type I interferon Rock bream Gene duplication Mx Type I interferon Iridovirus
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	Two type I interferon (IFN) genes, designated as rbIFN1 and rbIFN2, have been cloned and characterized in rock bream. They are both comprised of 5 exons and 4 introns, and are closely linked on the rock bream chromosome in a unique head-to-head configuration. Both genes encode 183 amino acid (aa) precursor with a putative 17 aa signal peptide in the N-terminal. Only one amino acid divergence is present between two IFNs. Compared with the type I IFNs in higher vertebrates, two rock bream IFNs possess conserved alpha helical structure and share approximately 20% identity in aa sequence. The highest aa sequence homology (83.2%) was found with European seabass IFNs. Phylogenetic analysis grouped two rock bream IFNs into the subgroup-d of two-cysteine containing IFNs. The gene synteny analysis revealed that they are orthologous with the zebrafish IFNφ4 on chromosome-12 and paralogous to each other, which are likely derived from a gene duplication event followed by an inversion. A number of cis-regulatory elements associated with immune response including 15 IRF and 6 NF-κB binding sites are predicted in the shared 4.5 kb 5’-flanking region. Highest constitutive expression of two IFNs was detected in blood cells and skin. Their expression in blood cells and head kidney was up-regulated by lipopolysaccharide, poly I:C, Edwardsiella tarda, Streptococcus iniae and iridovirus. Furthermore, recombinant rbIFN1 protein produced by E. coli induced a rapid and transient expression of the interferon inducible Mx gene in head kidney cells. These results suggest that two duplicated type I IFN genes are involved in rock bream host response to both viral and bacterial pathogens. ► Two duplicated type I interferon genes have been identified in rock bream. ► They belong to subgroup-d of two cysteine containing IFNs and are orthologous to zebrafish IFNφ4. ► The expression of two IFNs showed significant induction upon challenge with different immune stimuli in blood and head kidney. ► The rbIFN1 produced in E. coli showed bioactivity of inducing Mx gene expression. ► Two IFNs are involved in in rock bream host response to both viral and bacterial pathogens.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1050-4648 1095-9947
DOI:	10.1016/j.fsi.2012.07.018