Molecular cloning and functional characterization of two duplicated two-cysteine containing type I interferon genes in rock bream Oplegnathus fasciatus
- Authors
- Wan, Q., Wicramaarachchi, W.D., Whang, I., Lim, B.S., Oh, M.J., Jung, S.J., Kim, H.C., Yeo, S.Y., and Lee, J.
- ID
- ZDB-PUB-120815-19
- Date
- 2012
- Source
- Fish & shellfish immunology 33(4): 886-898 (Journal)
- Registered Authors
- Yeo, Sang-Yeob
- Keywords
- type I interferon, rock bream, gene duplication, iridovirus, Mx
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- Base Sequence
- Blood Cells/immunology
- Blood Cells/metabolism
- Chromosomes, Artificial, Bacterial/genetics
- Cloning, Molecular
- Cysteine/genetics
- Cysteine/immunology*
- DNA Virus Infections/immunology
- DNA Virus Infections/veterinary
- Edwardsiella tarda/physiology
- Enterobacteriaceae Infections/immunology
- Enterobacteriaceae Infections/veterinary
- Escherichia coli/genetics
- Fish Proteins/genetics
- Fish Proteins/immunology*
- GTP-Binding Proteins/chemistry
- GTP-Binding Proteins/genetics*
- Gene Duplication
- Genomic Library
- Head Kidney/immunology
- Head Kidney/metabolism
- Immunity, Innate
- Interferon Type I/genetics
- Interferon Type I/immunology*
- Iridoviridae/physiology
- Lipopolysaccharides/immunology
- Lipopolysaccharides/pharmacology
- Molecular Sequence Data
- Myxovirus Resistance Proteins
- Organ Specificity
- Perciformes/genetics
- Perciformes/immunology*
- 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
- PubMed
- 22889848 Full text @ Fish Shellfish Immunol.
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.