Characterisation of gamma-interferon responsive promoters in fish.

R. Castro, Samuel Allen Moore Martin, Steven Bird, J. Lamas, Christopher John Secombes

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Reporter constructs of three interferon (IFN)-gamma-induced rainbow trout genes were generated to examine specificity to type I or type II IFN. Constructs included gamma IP-10, LMP2 and TAP2 and were used to transfect trout fibroblast cells (RTG-2) which were then exposed to rainbow trout rIFNs. The gamma IP-10 construct showed high reporter activity even in the absence of rIFNs. The LMP2 promoter contained one GAS element and two double ISRE elements, of four constructs made, only those with ISRE elements showed significant reporter activity following rIFN-gamma stimulation. The TAP2 regulatory region contained two GAS, two ISRE and one C/EBP element from which four constructs were made. Reporter expression for the construct containing all five elements showed an 11- and 2-fold increase in response to rIFN-gamma and type I rIFN, respectively. Constructs containing only the GAS elements did not respond to rIFNs. The TAP2 construct with two ISRE and the C/EBP gave the greatest dose-dependent reporter response to rIFN-gamma, with no significant response to type I rIFN. These data suggest that the ISRE elements, or elements nearby, are essential for the induction of type II IFN responsive genes in trout. The TAP2 construct is a candidate to develop a IFN-gamma reporter stable cell line. (C) 2008 Elsevier Ltd. All rights reserved.

Original languageEnglish
Pages (from-to)3454-3462
Number of pages8
JournalMolecular Immunology
Volume45
Issue number12
Early online date5 May 2008
DOIs
Publication statusPublished - Jul 2008

Keywords

  • interferons
  • promoter
  • reporter constructs
  • ISRE
  • gas
  • trout oncorhynchus-mykiss
  • class-I pathway
  • rainbow-trout
  • IFN-Gamma
  • MX1 promoter
  • functional-characterization
  • reporter system
  • teleost fish
  • host-defense
  • LMP2 genes

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