Molecular and functional characterization of pigeon (Columba livia) tumor necrosis factor receptor-associated factor 3

Dev Comp Immunol. 2017 Apr:69:51-59. doi: 10.1016/j.dci.2016.12.005. Epub 2016 Dec 23.

Abstract

Tumor necrosis factor receptor-associated factor 3 (TRAF3) plays a key antiviral role by promoting type I interferon production. We cloned the pigeon TRAF3 gene (PiTRAF3) according to its predicted mRNA sequence to investigate its function. The 1704-bp full-length open reading frame encodes a 567-amino acid protein. One Ring finger, two TRAF-type Zinc fingers, one Coiled coil, and one MATH domain were inferred. RT-PCR showed that PiTRAF3 was expressed in all tissues, with relatively weak expression in the heart and liver. In HEK293T cells, over-expression of wild-type, △Ring, △Zinc finger, and △Coiled coil PiTRAF3, but not a △MATH form, significantly increased IFN-β promoter activity. Zinc finger and Coiled coil domains were essential for NF-κB activation. In chicken HD11 cells, PiTRAF3 increased IFN-β promoter activity and four domains were all contributing. R848 stimulation of pigeon peripheral blood mononuclear cells and splenocytes significantly increased expression of PiTRAF3 and the inflammatory cytokine genes CCL5, IL-8, and IL-10. These data demonstrate TRAF3's innate immune function and improve understanding of its involvement in poultry antiviral defense.

Keywords: Cloning; Function; Identification; Pigeon; Tumor necrosis factor receptor-associated factor 3.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Avian Proteins / genetics
  • Avian Proteins / metabolism*
  • Chickens
  • Cloning, Molecular
  • Columbidae / immunology*
  • HEK293 Cells
  • Humans
  • Immunity, Innate*
  • Inflammation / immunology*
  • Interferon-beta / genetics
  • NF-kappa B / metabolism
  • Promoter Regions, Genetic / genetics
  • Protein Conformation
  • Protein Domains / genetics
  • Protein Engineering
  • TNF Receptor-Associated Factor 3 / genetics
  • TNF Receptor-Associated Factor 3 / metabolism*
  • Transcriptional Activation

Substances

  • Avian Proteins
  • NF-kappa B
  • TNF Receptor-Associated Factor 3
  • Interferon-beta