Molecular cloning and characterization of crucian carp (Carassius auratus L.) interferon regulatory factor 7

Fish Shellfish Immunol. 2003 Nov;15(5):453-66. doi: 10.1016/s1050-4648(03)00025-1.

Abstract

Interferon (IFN) can induce an antiviral state via interferon-regulatory transcription factors (IRFs), which bind to and control genes directed by the interferon-stimulated response element (ISRE). Here we describe a fish IRF, termed CaIRF7, cloned from a subtractive cDNA library which is constructed with mRNAs obtained from crucian carp (Carassius auratus L.) blastulae embryonic (CAB) cells infected by UV-inactivated GCHV and mock-infected cells. CaIRF7 cDNA was found to be 1816 bp in length, with a 42 bp 5'UTR and a 508 bp 3'UTR. The open reading frame translates into 421 amino acids in which a DNA-binding domain (DBD) containing the repeated tryptophan motif and IRFs association domain have been identified. Like chicken GgIRF3, CaIRF7 was most similar to mammalian IRF7 with 27 to 30% identity overall and some 37% identity in their DBDs. A single transcript of 1.9 kb was detected in virally induced CAB cells by virtual Northern blotting. RT-PCR analysis revealed a wide tissue distribution of CaIRF7 constitutive expression, with detectable transcript in non-infected CAB cells and various tissues of healthy crucian carp. In addition, CaIRF7 expression was differentially increased by stimulation of the CAB cells with active GCHV, UV-inactivated GCHV or CAB IFN, indicating that the activation of CaIRF7 was directly regulated by IFN.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Blastula / immunology
  • Blotting, Northern
  • Cloning, Molecular*
  • Cluster Analysis
  • DNA Primers
  • DNA, Complementary / genetics
  • DNA-Binding Proteins / genetics*
  • DNA-Binding Proteins / immunology*
  • Gene Expression Profiling
  • Goldfish / genetics
  • Goldfish / immunology*
  • Interferon Regulatory Factor-7
  • Molecular Sequence Data
  • Phylogeny
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Alignment

Substances

  • DNA Primers
  • DNA, Complementary
  • DNA-Binding Proteins
  • Interferon Regulatory Factor-7

Associated data

  • GENBANK/AY177629