Molecular cloning and characterization of interferon regulatory factors 4 and 8 (IRF-4 and IRF-8) in rainbow trout, Oncorhynchus mykiss

Jason W Holland, Asma Karim, Tiehui Wang, Ayham Alnabulsi, Jamie Scott, Bertrand Collet, Muhammad S Mughal, Christopher J Secombes, Steve Bird (Corresponding Author)

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Mammalian interferon regulatory factor (IRF)4 (PIP, LSIRF, and ICSAT) and IRF8 (ICSBP) are known to be critical in regulating a spectrum of functional and developmental processes in lymphomyeloid cell lineages either through direct binding to IRF-E motifs in target gene promoters or indirectly by binding to composite motifs recognized by Ets family members, PU.I and Sp.I. Here we report, for the first time in fish, the sequencing and characterization of full-length cDNA homologues of rainbow trout (rt) IRF4 and rtIRF8. The rtIRF4 molecule consists of 1848 bp with a 45 bp 5' UTR and a predicted 378 bp 3' UTR translating into a 474 aa protein. RtIRF8 consists of 1951 bp with a 52 bp 5' UTR and a 564 bp 3' UTR translating into a 444 aa protein. Each gene possesses a putative DNA binding domain (DBD) containing the tryptophan pentad-repeat domain found in all IRF family members. Both molecules also possess a well conserved IRF association domain (IAD). The presence of these domains along with phylogenetic analysis places the two genes in the IRF4 subfamily. Both genes were detected in a range of trout tissues where IRF8 was the overall predominant transcript. Consistent with mammalian studies, the highest expression levels of IRF4 and IRF8 were observed in the lymphomyeloid-rich fish tissues, spleen, head kidney and gills. IRF8 expression in stimulated trout splenocytes was significantly up-regulated by polyinosinic:polycytidylic acid (poly I:C), trout recombinant (r)IL-15, phorbol 12-myristate 13-acetate (PMA), and phytohaemagglutinin (PHA) treatment whilst remaining refractory towards lipopolysaccharide (LPS) treatment. IRF4 was significantly down-regulated by LPS stimulation and remained refractory towards poly I:C, trout rIL15, and PHA. PMA stimulation elicited a significant upregulation of IRF4 expression. Overall, these data support the premise that these IRFs are likely to play important roles in the functional and developmental processes occurring in fish lymphomyeloid tissues.
Original languageEnglish
Pages (from-to)157-166
Number of pages10
JournalFish & Shellfish Immunology
Volume29
Issue number1
Early online date16 Mar 2010
DOIs
Publication statusPublished - Jul 2010

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Keywords

  • rainbow tout
  • interferon regulatory factor
  • IRF4
  • IRF8
  • fish
  • lymphoid development
  • trout splenocytes
  • gene regulation
  • gene upregulation

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