title: Molecular and cellular mechanisms underlying iron transport deficiency in microcytic anemia.
abstract: A mutation of the iron transporter Nramp2 (DMT1, Slc11a2) causes microcytic anemia in mk mice and in Belgrade rats by impairing iron absorption in the duodenum and in erythroid cells, causing severe iron deficiency. Both mk and Belgrade animals display a glycine-to-arginine substitution at position 185 (G185R) in the fourth predicted transmembrane domain of Nramp2. To study the molecular basis for the loss of function of Nramp2(G185R), we established cell lines stably expressing extracellularly tagged versions of wild-type (WT) or mutated transporters. Like WT Nramp2, the G185R mutant was able to reach the plasmalemma and endosomal compartments, but with reduced efficiency. Instead, a large fraction of Nramp2(G185R) was detected in the endoplasmic reticulum, where it was unstable and was rapidly degraded by a proteasome-dependent mechanism. Moreover, the stability of the mutant protein that reached the plasma membrane was greatly reduced, further diminishing its surface density at steady state. Last, the specific metal transport activity of plasmalemmal Nramp2(G185R) was found to be significantly depressed, compared with its WT counterpart. Thus, a singlepoint mutation results in multiple biosynthetic and functional defects that combine to produce the impaired iron deficiency that results in microcytic anemia.
authors.affiliation: Programme in Cell Biology, The Hospital for Sick Children, 555 University Avenue, Toronto, Ontario, Canada M5G 1X8. sga@sickkids.ca
authors.forename: Nicolas
authors.initials: N
authors.lastname: Touret
authors.forename: Natalia
authors.initials: N
authors.lastname: Martin-Orozco
authors.forename: Paul
authors.initials: P
authors.lastname: Paroutis
authors.forename: Wendy
authors.initials: W
authors.lastname: Furuya
authors.forename: Steven
authors.initials: S
authors.lastname: Lam-Yuk-Tseung
authors.forename: John
authors.initials: J
authors.lastname: Forbes
authors.forename: Philippe
authors.initials: P
authors.lastname: Gros
authors.forename: Sergio
authors.initials: S
authors.lastname: Grinstein
mesh_terms: D000740:Anemia / Q000235:genetics* / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport / Q000502:physiology; D016466:CHO Cells; D027682:Cation Transport Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D003546:Cysteine Endopeptidases / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D007501:Iron / Q000378:metabolism*; D033862:Iron-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D018374:LLC-PK1 Cells; D008247:Lysosomes / Q000378:metabolism; D009097:Multienzyme Complexes / Q000378:metabolism; D016296:Mutagenesis; D017354:Point Mutation; D046988:Proteasome Endopeptidase Complex; D017510:Protein Folding; D013552:Swine; D000099236:Solute Carrier Family 11, Member 2
chemical_list: D027682:Cation Transport Proteins; D003546:Cysteine Endopeptidases; D007501:Iron; D033862:Iron-Binding Proteins; D009097:Multienzyme Complexes; D046988:Proteasome Endopeptidase Complex; D000099236:Solute Carrier Family 11, Member 2