Human mitochondrial ferritin expressed in HeLa cells incorporates iron and affects cellular iron metabolism.
Mitochondrial ferritin (MtF) is a newly identified ferritin encoded by an intronless gene on chromosome 5q23.1. The mature recombinant MtF has a ferroxidase center and binds iron in vitro similarly to H-ferritin. To explore the structural and functional aspects of MtF, we expressed the following forms in HeLa cells: the MtF precursor (approximately 28 kDa), a mutant MtF precursor with a mutated ferroxidase center, a truncated MtF lacking the approximately 6-kDa mitochondrial leader sequence, and a chimeric H-ferritin with this leader sequence. The experiments show that all constructs with the leader sequence were processed into approximately 22-kDa subunits that assembled into multimeric shells electrophoretically distinct from the cytosolic ferritins. Mature MtF was found in the matrix of mitochondria, where it is a homopolymer. The wild type MtF and the mitochondrially targeted H-ferritin both incorporated the (55)Fe label in vivo. The mutant MtF with an inactivated ferroxidase center did not take up iron, nor did the truncated MtF expressed transiently in cytoplasm. Increased levels of MtF both in transient and in stable transfectants resulted in a greater retention of iron as MtF in mitochondria, a decrease in the levels of cytosolic ferritins, and up-regulation of transferrin receptor. Neither effect occurred with the mutant MtF with the inactivated ferroxidase center. Our results indicate that exogenous iron is as available to mitochondrial ferritin as it is to cytosolic ferritins and that the level of MtF expression may have profound consequences for cellular iron homeostasis.
Corsi
Barbara
B
Section of Chemistry, Faculty of Medicine, University of Brescia, Brescia, 25100 Italy.
Cozzi
Anna
A
Arosio
Paolo
P
Drysdale
Jim
J
Santambrogio
Paolo
P
Campanella
Alessandro
A
Biasiotto
Giorgio
G
Albertini
Alberto
A
Levi
Sonia
S
D015153:Blotting, Western; D002460:Cell Line; D003600:Cytosol / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005293:Ferritins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007438:Introns; D007501:Iron / Q000737:chemistry / Q000378:metabolism*; D000090463:Iron Deficiencies; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000378:metabolism*; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding; D011990:Receptors, Transferrin / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D015854:Up-Regulation
D011990:Receptors, Transferrin; D005293:Ferritins; D007501:Iron