Molecular and expression analysis of an interferon-gamma-inducible guanylate-binding protein from rainbow trout ( Oncorhynchus mykiss)

Abstract

Guanylate-binding proteins (GBPs) are some of the most abundant proteins accumulating in mammalian cells in response to interferon- γ (IFN- γ). GBPs have been suggested to function in antiviral activity, macrophage activation, fibroblast proliferation and inhibition of endothelial cell proliferation and invasiveness. Here we confirm that IFN- γ-inducible GBP also exist in fish. A 2 kb GBP cDNA was cloned from head kidney of rainbow trout treated with an IFN-inducing compound. The open reading frame predicts a 635 amino acid protein (rbtGBP) of 72.7 kDa possessing a tripartite GTP binding motif and a secondary structure similar to human GBP1. Like most mammalian GBPs, rbtGBP possesses an isoprenylation motif at the C-terminal end. The overall amino acid sequence identity between rbtGBP and mammalian GBPs is only 41–47%, however. The rainbow trout macrophage cell line RTS11 showed a dose-dependent increase in rbtGBP transcripts in response to IFN- γ after 6 h of stimulation, with rbtGBP being undetectable in non-treated RTS11 cells. Moreover, polyinosinic polycytidylic acid (poly I:C) induced increased GBP transcript levels in RTS11 and RTG2 cells after 4–6 h of stimulation, and in head kidney and liver of live fish after 24 h. These studies suggest that rbtGBP is an early response gene in rainbow trout, which may have similar functions in IFN- γ mediated responses as mammalian GBPs.