Biological characterization, expression, and functional analysis of tumor necrosis factor receptor-associated factor 6 in Nile tilapia (Oreochromis niloticus)

Fish Shellfish Immunol. 2018 Sep:80:497-504. doi: 10.1016/j.fsi.2018.06.036. Epub 2018 Jun 19.

Abstract

Nile tilapia (Oreochromis niloticus) is a pivotal economic fish that has been plagued by Streptococcus agalactiae infections for many years. Tumor necrosis factor receptor-associated factor 6 (TRAF6) is a crucial adaptor molecule of the interleukin-1 receptor/Toll-like receptor (IL-1/TLR) superfamily, which can trigger downstream signaling cascades involved in innate immunity. In this study, the full-length cDNA of TRAF6 was cloned from O. niloticus (named On-TRAF6), which has an open reading frame of 1716 bp, and encode a polypeptide of 571 amino acids. The predicted amino acid sequence of On-TRAF6 contained the characteristic motifs of TRAF proteins, including a Zinc finger of RING-type, two Zinc fingers of TRAF-type, and a MATH (meprin and TRAF homology) domain. Multiple sequence alignment revealed that On-TRAF6 shares a relatively high level of identity with those of other fishes (64-98%). In healthy tilapia, mRNA expression of On-TRAF6 could be detected in all the examined tissues and the highest expression occurred in the spleen. Moreover, we found that On-TRAF6 was involved immune response of Nile tilapia following the stimulation with Streptococcus agalactiae and polyinosinic: polycytidylic acid (Poly I:C) when determined by using qPCR. The result of subcellular localization showed that On-TRAF6 distributed in the cytoplasm, and over-expression of On-TRAF6 could strongly activated NF-кB pathway in HEK293T cells. These findings indicated that On-TRAF6 may play an important role in the immune response to intracellular bacteria in Nile tilapia.

Keywords: Infection; Oreochromis niloticus; TRAF6; mRNA expression.

MeSH terms

  • Animals
  • Cichlids / genetics*
  • Cichlids / immunology
  • Fish Proteins / genetics*
  • Fish Proteins / immunology
  • Fish Proteins / metabolism
  • HEK293 Cells
  • Head Kidney / immunology
  • Humans
  • Intestines / immunology
  • NF-kappa B / metabolism
  • Poly I-C / pharmacology
  • Spleen / immunology
  • Streptococcus agalactiae / immunology
  • TNF Receptor-Associated Factor 6 / genetics*
  • TNF Receptor-Associated Factor 6 / immunology
  • TNF Receptor-Associated Factor 6 / metabolism

Substances

  • Fish Proteins
  • NF-kappa B
  • TNF Receptor-Associated Factor 6
  • Poly I-C