Sequence and Expression Analysis of Interferon Regulatory Factor 10 (IRF10) in Three Diverse Teleost Fish Reveals Its Role in Antiviral Defense

Background Interferon regulatory factor (IRF) 10 was first found in birds and is present in the genome of other tetrapods (but not humans and mice), as well as in teleost fish. The functional role of IRF10 in vertebrate immunity is relatively unknown compared to IRF1-9. The target of this research was to clone and characterize the IRF10 genes in three economically important fish species that will facilitate future evaluation of this molecule in fish innate and adaptive immunity. Molecular Characterization of IRF10 in Three Fish Species In the present study, a single IRF10 gene was cloned in grass carp Ctenopharyngodon idella and Asian swamp eel Monopterus albus, and two, named IRF10a and IRF10b, in rainbow trout Oncorhynchus mykiss. The fish IRF10 molecules share highest identities to other vertebrate IRF10s, and have a well conserved DNA binding domain, IRF-associated domain, and an 8 exon/7 intron structure with conserved intron phase. The presence of an upstream ATG or open reading frame (ORF) in the 5’-untranslated region of different fish IRF10 cDNA sequences suggests potential regulation at the translational level, and this has been verified by in vitro transcription/translation experiments of the trout IRF10a cDNA, but would still need to be validated in fish cells. Expression Analysis of IRF10 In Vivo and In Vitro Both trout IRF10 paralogues are highly expressed in thymus, blood and spleen but are relatively low in head kidney and caudal kidney. Trout IRF10b expression is significantly higher than IRF10a in integumentary tissues i.e. gills, scales, skin, intestine, adipose fin and tail fins, suggesting that IRF10b may be more important in mucosal immunity. The expression of both trout IRF10 paralogues is up-regulated by recombinant IFN-γ. The expression of the IRF10 genes is highly induced by Poly I:C in vitro and in vivo, and by viral infection, but is less responsive to peptidoglycan and bacterial infection, suggesting an important role of fish IRF10 in antiviral defense.

[1]  Samuel A. M. Martin,et al.  Two copies of the genes encoding the subunits of putative interleukin (IL)-4/IL-13 receptors, IL-4Rα, IL-13Rα1 and IL-13Rα2, have been identified in rainbow trout (Oncorhynchus mykiss) and have complex patterns of expression and modulation , 2011, Immunogenetics.

[2]  Klaus Wethmar,et al.  Upstream open reading frames: Molecular switches in (patho)physiology , 2010, BioEssays : news and reviews in molecular, cellular and developmental biology.

[3]  Peer Bork,et al.  SMART 7: recent updates to the protein domain annotation resource , 2011, Nucleic Acids Res..

[4]  Tiehui Wang,et al.  Supporting Information for DOI 10.1002/eji.201344273 , 2014 .

[5]  T. Taniguchi,et al.  The IRF family transcription factors in immunity and oncogenesis. , 2008, Annual review of immunology.

[6]  Samuel A. M. Martin,et al.  Functional Characterization of a Nonmammalian IL-21: Rainbow Trout Oncorhynchus mykiss IL-21 Upregulates the Expression of the Th Cell Signature Cytokines IFN-γ, IL-10, and IL-22 , 2011, The Journal of Immunology.

[7]  G. Scapigliati,et al.  The production and bioactivity of rainbow trout (Oncorhynchus mykiss) recombinant IL-1 beta. , 2001, Veterinary immunology and immunopathology.

[8]  P. Nie,et al.  The gene and virus-induced expression of IRF-5 in grass carp Ctenopharyngodon idella. , 2010, Veterinary immunology and immunopathology.

[9]  Tiehui Wang,et al.  The expanding repertoire of the IL-12 cytokine family in teleost fish: Identification of three paralogues each of the p35 and p40 genes in salmonids, and comparative analysis of their expression and modulation in Atlantic salmon Salmo salar. , 2014, Developmental and comparative immunology.

[10]  J. Meseguer,et al.  Post-transcriptional regulation of cytokine genes in fish: A role for conserved AU-rich elements located in the 3'-untranslated region of their mRNAs. , 2007, Molecular immunology.

[11]  B. Collet Innate immune responses of salmonid fish to viral infections. , 2014, Developmental and comparative immunology.

[12]  Thomas L. Madden,et al.  Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. , 1997, Nucleic acids research.

[13]  Koichiro Tamura,et al.  MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. , 2013, Molecular biology and evolution.

[14]  K. Handberg,et al.  Transcriptional profiles of chicken embryo cell cultures following infection with infectious bursal disease virus , 2006, Archives of Virology.

[15]  N. J. Olesen,et al.  Viral haemorrhagic septicaemia virus in marine fish and its implications for fish farming--a review. , 2005, Journal of fish diseases.

[16]  N. Roher,et al.  Peptidoglycan, not endotoxin, is the key mediator of cytokine gene expression induced in rainbow trout macrophages by crude LPS. , 2010, Molecular immunology.

[17]  Ledion Bitincka,et al.  MatGAT: An application that generates similarity/identity matrices using protein or DNA sequences , 2003, BMC Bioinformatics.

[18]  P. Nie,et al.  Open Access Research Article , 2022 .

[19]  Yongan Zhang,et al.  IFN Regulatory Factor 10 Is a Negative Regulator of the IFN Responses in Fish , 2014, The Journal of Immunology.

[20]  R D Appel,et al.  Protein identification and analysis tools in the ExPASy server. , 1999, Methods in molecular biology.

[21]  K. Khabar Cellular and Molecular Life Sciences REVIEW Post-transcriptional control during chronic inflammation , 2022 .

[22]  C. Secombes,et al.  Sequencing of a second interleukin-10 gene in rainbow trout Oncorhynchus mykiss and comparative investigation of the expression and modulation of the paralogues in vitro and in vivo. , 2011, Fish & shellfish immunology.

[23]  C. Secombes,et al.  Four CISH paralogues are present in rainbow trout Oncorhynchus mykiss: differential expression and modulation during immune responses and development. , 2014, Molecular immunology.

[24]  J. Burnside,et al.  A Novel Interferon Regulatory Factor (IRF), IRF-10, Has a Unique Role in Immune Defense and Is Induced by the v-Rel Oncoprotein , 2002, Molecular and Cellular Biology.

[25]  G. Zhu,et al.  Advances in research of fish immune-relevant genes: a comparative overview of innate and adaptive immunity in teleosts. , 2013, Developmental and comparative immunology.

[26]  R. Guyomard,et al.  An Immune-Related Gene Evolved into the Master Sex-Determining Gene in Rainbow Trout, Oncorhynchus mykiss , 2012, Current Biology.

[27]  M. Cáccamo,et al.  Conservation and divergence of gene families encoding components of innate immune response systems in zebrafish , 2007, Genome Biology.

[28]  Qi-ya Zhang,et al.  Identification and characterization of interferon regulatory factor-1 from orange-spotted grouper (Epinephelus coioides) , 2010, Molecular Biology Reports.

[29]  C. Secombes,et al.  Identifying potential virulence determinants in viral haemorrhagic septicaemia virus (VHSV) for rainbow trout. , 2009, Diseases of aquatic organisms.

[30]  Rodrigo Lopez,et al.  Multiple sequence alignment with the Clustal series of programs , 2003, Nucleic Acids Res..

[31]  R. Hrdličková,et al.  Dynamic evolution of immune system regulators: the history of the interferon regulatory factor family. , 2009, Molecular biology and evolution.

[32]  O. A. Volkova,et al.  uORFs, reinitiation and alternative translation start sites in human mRNAs , 2008, FEBS letters.

[33]  Oie Diagnostic manual for aquatic animal diseases , 2017 .

[34]  C. Watson,et al.  Diet of the Nonindigenous Asian Swamp Eel in Tropical Ornamental Aquaculture Ponds in West-Central Florida , 2007 .

[35]  L. Jun,et al.  A detection method for grass carp hemorrhagic virus (GCHV) based on a reverse transcription polymerase chain reaction , 1997 .

[36]  D. R. Jones,et al.  Characterisation of mucosal and systemic immune responses in rainbow trout (Oncorhynchus mykiss) using surface plasmon resonance. , 2000, Fish & shellfish immunology.

[37]  W. Royer,et al.  Structural insights into interferon regulatory factor activation. , 2010, Cellular signalling.

[38]  Pin Nie,et al.  Global characterization of interferon regulatory factor (IRF) genes in vertebrates: Glimpse of the diversification in evolution , 2010, BMC Immunology.

[39]  Ronggai Li,et al.  Sequence and expression analysis of rainbow trout CXCR2, CXCR3a and CXCR3b aids interpretation of lineage-specific conversion, loss and expansion of these receptors during vertebrate evolution , 2014, Developmental and comparative immunology.

[40]  C. Secombes,et al.  Cloning of the IL-1β3 gene and IL-1β4 pseudogene in salmonids uncovers a second type of IL-1β gene in teleost fish. , 2012, Developmental and comparative immunology.

[41]  Ronggai Li,et al.  Two Types of TNF-α Exist in Teleost Fish: Phylogeny, Expression, and Bioactivity Analysis of Type-II TNF-α3 in Rainbow Trout Oncorhynchus mykiss , 2013, The Journal of Immunology.

[42]  C. Secombes,et al.  Complete sequencing and expression of three complement components, C1r, C4 and C1 inhibitor, of the classical activation pathway of the complement system in rainbow trout Oncorhynchus mykiss , 2003, Immunogenetics.

[43]  C. Secombes,et al.  Bioactivity studies of rainbow trout (Oncorhynchus mykiss) interleukin-6: effects on macrophage growth and antimicrobial peptide gene expression. , 2011, Molecular immunology.

[44]  T. Taniguchi,et al.  High-mobility group box family of proteins: ligand and sensor for innate immunity. , 2012, Trends in immunology.

[45]  T. Taniguchi,et al.  IRF family of transcription factors as regulators of host defense. , 2001, Annual review of immunology.

[46]  I. Hirono,et al.  Molecular cloning and expression analysis of interferon regulatory factor 10 (IRF10) in Japanese flounder, Paralichthys olivaceus. , 2011, Fish & shellfish immunology.