Molecular cloning and multifunctional characterization of GRIM-19 (gene associated with retinoid-interferon-induced mortality 19) homologue from turbot (Scophthalmus maximus)

Dev Comp Immunol. 2014 Mar;43(1):96-105. doi: 10.1016/j.dci.2013.11.004. Epub 2013 Nov 13.

Abstract

GRIM-19 (gene associated with retinoid-interferon-induced mortality 19), a novel cell death regulatory gene, plays important roles in cell apoptosis, embryogenesis, mitochondrial respiratory chain and immune response. To date, little information is known about fish GRIM-19 characteristics except orange-spotted grouper (Epinephelus coioides). Here a new GRIM-19 gene is identified and characterized from turbot (Scophthalmus maximus), an economic marine fish in China and Europe. Briefly, turbot GRIM-19 is a 595-bp gene encoding a 144 amino acids protein, which shares the closest relationship with Atlantic halibut (Hippoglossus hippoglossus). The expression of turbot grim-19 in liver, spleen and kidney is up-regulated by the infection of Vibrio anguillarum and LCDV (lymphocystis disease virus). Subsequently, a recombinant protein of turbot GRIM-19 is acquired and the anti-bacterial function is proved by liquid culture inhibition experiment. The subcellular location indicates that turbot GRIM-19 is co-localized with STAT3 in the cytoplasm, which is mainly determined by GRIM-19 41-84 amino acids and STAT3 1-321 amino acids. Finally, the involvements of turbot GRIM-19 in cell apoptosis and NF-κB pathway are investigated. All these data help to understand GRIM-19 function in fish, as well as provide the application possibility of GRIM-19 in fish disease resistance breeding.

Keywords: Cell apoptosis; GRIM-19; Immune response; NF-κB; STAT3; Turbot (Scophthalmus maximus).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Apoptosis Regulatory Proteins / genetics
  • Apoptosis Regulatory Proteins / isolation & purification
  • Apoptosis Regulatory Proteins / metabolism*
  • Cells, Cultured
  • Cloning, Molecular
  • DNA Virus Infections / immunology*
  • Fish Proteins / genetics
  • Fish Proteins / isolation & purification
  • Fish Proteins / metabolism*
  • Flatfishes / immunology*
  • Flounder / immunology
  • Humans
  • Iridoviridae / immunology*
  • Molecular Sequence Data
  • NADH, NADPH Oxidoreductases / genetics
  • NF-kappa B / metabolism
  • STAT3 Transcription Factor / metabolism
  • Sequence Homology, Amino Acid
  • Signal Transduction
  • Transgenes / genetics
  • Up-Regulation
  • Vibrio / immunology*
  • Vibrio Infections / immunology*

Substances

  • Apoptosis Regulatory Proteins
  • Fish Proteins
  • NF-kappa B
  • STAT3 Transcription Factor
  • NADH, NADPH Oxidoreductases
  • NDUFA13 protein, human