Characterization of the iron transporter DMT1 (NRAMP2/DCT1) in red blood cells of normal and anemic mk/mk mice.
Divalent metal transporter 1 (DMT1) is the major transferrin-independent iron uptake system at the apical pole of intestinal cells, but it may also transport iron across the membrane of acidified endosomes in peripheral tissues. Iron transport and expression of the 2 isoforms of DMT1 was studied in erythroid cells that consume large quantities of iron for biosynthesis of hemoglobin. In mk/mk mice that express a loss-of-function mutant variant of DMT1, reticulocytes have a decreased cellular iron uptake and iron incorporation into heme. Interestingly, iron release from transferrin inside the endosome is normal in mk/mk reticulocytes, suggesting a subsequent defect in Fe(++) transport across the endosomal membrane. Studies by immunoblotting using membrane fractions from peripheral blood or spleen from normal mice where reticulocytosis was induced by erythropoietin (EPO) or phenylhydrazine (PHZ) treatment suggest that DMT1 is coexpressed with transferrin receptor (TfR) in erythroid cells. Coexpression of DMT1 and TfR in reticulocytes was also detected by double immunofluorescence and confocal microscopy. Experiments with isoform-specific anti-DMT1 antiserum strongly suggest that it is the non-iron-response element containing isoform II of DMT1 that is predominantly expressed by the erythroid cells. As opposed to wild-type reticulocytes, mk/mk reticulocytes express little if any DMT1, despite robust expression of TfR, suggesting a possible effect of the mutation on stability and targeting of DMT1 isoform II in these cells. Together, these results provide further evidence that DMT1 plays a central role in iron acquisition via the transferrin cycle in erythroid cells.
Canonne-Hergaux
F
F
Department of Biochemistry, McGill University, Montreal, QC, Canada.
Zhang
A S
AS
Ponka
P
P
Gros
P
P
D000740:Anemia / Q000097:blood*; D000818:Animals; D001692:Biological Transport / Q000235:genetics; D016466:CHO Cells; D027682:Cation Transport Proteins / Q000097:blood* / Q000235:genetics / Q000502:physiology; D006224:Cricetinae; D011992:Endosomes / Q000737:chemistry / Q000378:metabolism; D004910:Erythrocyte Membrane / Q000378:metabolism; D004912:Erythrocytes / Q000737:chemistry* / Q000648:ultrastructure; D015672:Erythroid Precursor Cells / Q000737:chemistry / Q000648:ultrastructure; D004921:Erythropoietin / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D006418:Heme / Q000378:metabolism; D006454:Hemoglobins / Q000096:biosynthesis; D006720:Homozygote; D007501:Iron / Q000097:blood; D033862:Iron-Binding Proteins; D004915:Leukemia, Erythroblastic, Acute; D051379:Mice; D008817:Mice, Mutant Strains; D018613:Microscopy, Confocal; D009154:Mutation; D010659:Phenylhydrazines / Q000494:pharmacology; D020033:Protein Isoforms / Q000235:genetics; D017701:Reticulocyte Count; D012156:Reticulocytes / Q000187:drug effects / Q000378:metabolism; D013154:Spleen / Q000187:drug effects / Q000378:metabolism; D014168:Transferrin / Q000378:metabolism; D014407:Tumor Cells, Cultured; D000099236:Solute Carrier Family 11, Member 2
D027682:Cation Transport Proteins; D004921:Erythropoietin; D006418:Heme; D006454:Hemoglobins; D007501:Iron; D033862:Iron-Binding Proteins; D010659:Phenylhydrazines; D020033:Protein Isoforms; D014168:Transferrin; D000099236:Solute Carrier Family 11, Member 2; C030299:phenylhydrazine