Isolation and characterization of channel catfish natural resistance associated macrophage protein gene

Natural resistance associated macrophage protein 1 (Nramp1) affects the ability of macrophages to kill pathogens. We cloned Nramp cDNA of channel catfish to identify potential molecular markers for disease resistance. Three different Nramp transcripts were identified: NrampCa-2912 nucleotides (nt), NrampCb-3245 nt, and NrampCc-3721 nt. At the 5' end, the transcripts have a common 2263 nt sequence containing the open reading frame. The differences are in the 3' untranslated region resulting from alternative splicing and polyadenylation. NrampCc is the predominant form expressed. The deduced 550 amino acid sequence of the channel catfish Nramp (NrampC) has high homology to Nramp from other vertebrates and a predicted conserved structure. The NrampC contains the 12 transmembrane domains, and the consensus transport motif. Post-transcriptional processing is also conserved. Phylogenetic analysis grouped NrampC with other fish Nramps and closer to Nramp2 than to Nramp1 of mammals. However, the catfish transcript does not contain an iron-responsive regulatory-protein binding site, a characteristic of Nramp2, and, like Nramp1, NrampC expression is induced in macrophage-rich tissues after exposure to lipopolysaccharide and in a macrophage cell line when stimulated. Thus NrampC is structurally closer to mammalian Nramp2 but may function similar to Nramp1.