Overexpression of the hereditary hemochromatosis protein, HFE, in HeLa cells induces and iron-deficient phenotype.
A transfectant HeLa cell clone expressing HFE under the control of a tetracycline-repressible promoter was generated. HFE expression was fully repressed by the presence of doxycycline, while it was strongly induced by growth in the absence of doxycycline. HFE accumulation was accompanied by a large (approximately 10-fold) decrease in H- and L-ferritin levels, by a approximately 3-4-fold increase in transferrin receptor, and a approximately 2-fold increase in iron regulatory protein activity. These indices of cellular iron deficiency were reversed by iron supplementation complexes. The overexpressed HFE immunoprecipitated together with transferrin receptor, indicating a physical association which is the likely cause for the observed approximately 30% decrease in 55Fe-transferrin incorporation after 18 h incubation. In the HFE-expressing cells the reduction in transferrin-mediated iron incorporation was partially compensated by a approximately 30% increase in non-transferrin iron incorporation from 55Fe-NTA, evident after prolonged, 18 h, incubations. The findings indicate that HFE binding to transferrin receptor reduces cellular iron availability and regulates the balance between transferrin-mediated and non-transferrin-mediated cellular iron incorporation.
Corsi
B
B
Dibit, Department of Biological and Technological Research, IRCCS H. San Raffaele, Milan, Italy.
Levi
S
S
Cozzi
A
A
Corti
A
A
Altimare
D
D
Albertini
A
A
Arosio
P
P
D001052:Apoferritins; D004318:Doxycycline / Q000494:pharmacology; D005293:Ferritins / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006680:HLA Antigens / Q000096:biosynthesis* / Q000235:genetics; D006367:HeLa Cells; D006432:Hemochromatosis / Q000235:genetics; D000071020:Hemochromatosis Protein; D015395:Histocompatibility Antigens Class I / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D000090463:Iron Deficiencies; D035925:Iron-Regulatory Proteins; D007506:Iron-Sulfur Proteins / Q000378:metabolism; D008565:Membrane Proteins; D011401:Promoter Regions, Genetic; D016601:RNA-Binding Proteins / Q000378:metabolism; D011990:Receptors, Transferrin / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid; D014162:Transfection
C103673:HFE protein, human; D006680:HLA Antigens; D000071020:Hemochromatosis Protein; D015395:Histocompatibility Antigens Class I; D035925:Iron-Regulatory Proteins; D007506:Iron-Sulfur Proteins; D008565:Membrane Proteins; D016601:RNA-Binding Proteins; D011990:Receptors, Transferrin; D005293:Ferritins; D001052:Apoferritins; D004318:Doxycycline