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Molecular cloning and functional characterization of two duplicated two-cysteine containing type I interferon genes in rock bream Oplegnathus fasciatus.
- Source :
-
Fish & shellfish immunology [Fish Shellfish Immunol] 2012 Oct; Vol. 33 (4), pp. 886-98. Date of Electronic Publication: 2012 Aug 07. - Publication Year :
- 2012
-
Abstract
- 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.<br /> (Copyright © 2012 Elsevier Ltd. All rights reserved.)
- Subjects :
- Amino Acid Sequence
Animals
Base Sequence
Blood Cells immunology
Blood Cells metabolism
Chromosomes, Artificial, Bacterial genetics
Cloning, Molecular
Cysteine genetics
DNA Virus Infections immunology
DNA Virus Infections veterinary
Edwardsiella tarda physiology
Enterobacteriaceae Infections immunology
Enterobacteriaceae Infections veterinary
Escherichia coli genetics
Fish Proteins genetics
GTP-Binding Proteins chemistry
Gene Duplication
Genomic Library
Head Kidney immunology
Head Kidney metabolism
Immunity, Innate
Interferon Type I genetics
Iridoviridae physiology
Lipopolysaccharides immunology
Lipopolysaccharides pharmacology
Molecular Sequence Data
Myxovirus Resistance Proteins
Organ Specificity
Perciformes genetics
Phylogeny
Poly I-C immunology
Real-Time Polymerase Chain Reaction veterinary
Reverse Transcriptase Polymerase Chain Reaction veterinary
Sequence Alignment veterinary
Sequence Homology, Amino Acid
Streptococcal Infections immunology
Streptococcal Infections veterinary
Streptococcus physiology
Cysteine immunology
Fish Proteins immunology
GTP-Binding Proteins genetics
Interferon Type I immunology
Perciformes immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1095-9947
- Volume :
- 33
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Fish & shellfish immunology
- Publication Type :
- Academic Journal
- Accession number :
- 22889848
- Full Text :
- https://doi.org/10.1016/j.fsi.2012.07.018