Functional characterization of iron transporter from Dictyostelium discoideum as a model of cellular iron homeostasis

Iron plays a central role in a large number of essential cellular functions but it is also potentially toxic being able to generate reactive oxygen species. The social amoeba Dictyostelium discoideum possesses several iron genes with the exception of transferrin, ferritin and TFR (Bozzaro et al., 2013; Peracino et al., 2013) and represents a model for the study of cellular iron homeostasis showing subcellular localization of iron transporters resembling that of macrophages. In particular, D. discoideum expresses the ortholog of Nramp1 transporter in phago-lysosomes and that of Nramp2 in the contractile vacuole. To better understand the function of dd Nramp1, the protein was expressed in Xenopus laevis oocytes by cRNA injection and functionally tested by radiochemical and electrophysiological techniques. To increase the surface localization of dd Nramp1, its N and C termini were replaced with the corresponding regions of the murine DMT1, which shows a high level of expression in the membrane of X. laevis oocytes (Gunshin et al., 1997). Dd Nramp1 is an electrogenic proton-dependent divalent metal ion transporter with a cation selectivity comparable to that of the murine DMT1 (Illing et al., 2012). It transports ferrous but not ferric iron and it is partially inhibited by Na+.