Mutations of FLVCR1 in posterior column ataxia and retinitis pigmentosa result in the loss of heme export activity.

The feline leukemia virus subgroup C receptor 1 (FLVCR1) is a heme exporter that maintains the intracellular heme concentration. FLVCR1 was previously assumed to be involved in Diamond-Blackfan anemia, and it was recently reported that mutations in the FLVCR1 gene are found in patients with posterior column ataxia and retinitis pigmentosa (PCARP). Four mutations in FLVCR1 (Asn121Asp, Cys192Arg, Ala241Thr, and Gly493Arg) are located within putative transmembrane domains; however, the effects of FLVCR1 mutations on PCARP are unclear. In this study, we analyzed the function of FLVCR1 mutants by using a fluorescent heme analog as a transporter substrate, and found that all 4 FLVCR1 mutants lost their heme export activity. To investigate the mechanism responsible for this loss of activity, we determined the subcellular localization of FLVCR1 mutants. FLVCR1 mutants did not localize to the plasma membrane and were observed in intracellular structures, including lysosomes. We hypothesize that the loss of function of FLVCR1 mutants is caused by their mislocation. We examined the half-life of FLVCR1 in cells, which was >16h for wild-type FLVCR1 compared with 2-4h for the mutants. Based on these results, we propose that FLVCR1 mutants failed to fold properly in the ER, were rapidly degraded in the lysosomes, and therefore, could not export heme out of cells. Thus, accumulation of heme in FLVCR1-mutant cells could cause cellular toxicity.