Functional characterization of an immunity-related GTPase gene in immune defense from obscure puffer (Takifugu obscurus)

Immunity-related GTPases (IRGs) are a family of large interferon-inducible GTPases that function in effective host defense against invading pathogens. IRGs have been extensively studied in mammals for their roles in the elimination of intracellular pathogens; however, their homologs in lower vertebrates are not well known. In this study, an IRG from obscure puffer (Takifugu obscurus), ToIRG, was identified and further characterized for its functional activity. The ToIRG gene encodes a protein of 396 amino acids containing a typical N-terminal GTPase domain with three conserved motifs. Phylogenetic analysis revealed that it has a closer evolutionary relationship with mammalian GKS IRGs. Gene expression profile analysis revealed that ToIRG was ubiquitously expressed in all tested healthy tissues of obscure puffer and upregulated in response to Aeromonas hydrophila or Edwardsiella tarda challenge. The subcellular localization of ToIRG is characterized as condensed forms around the nucleus. Importantly, an antimicrobial assay in vitro suggested that ToIRG enhanced the ability of host cells to resist both intracellular (E. tarda) and extracellular pathogens (A. hydrophila). Taken together, these results provide the functional characterization of obscure puffer IRGs in immune defense, which is the first study to reveal the function of IRGs in bony fish and will provide important insights into the evolutionary divergence of IRGs. •The immunity-related GTPase from obscure puffer (ToIRG) was identified.•ToIRG shared highly similar molecular features with mammalian GKS IRGs.•ToIRG was dramatically upregulated after Aeromonas hydrophila or Edwardsiella tarda infection.•ToIRG is localized in the cytoplasm and characterized as condensed form around the nucleus.•ToIRG helped host cell to resist against both intracellular (E.tarda) and extracellular pathogens (A.hydrophila).