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Apical distribution of HFE–β2-microglobulin is associated with inhibition of apical iron uptake in intestinal epithelia cells

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Abstract

Mutations in the HFE gene result in hereditary hemochromatosis, a disorder of iron metabolism characterized by increased intestinal iron absorption. Based on the observation that ectopic expression of HFE strongly inhibits apical iron uptake (Arredondo et al., 2001, FASEB J 15, 1276–1278), a negative regulation of HFE on the apical membrane transporter DMT1 was proposed as a mechanism by which HFE regulates iron absorption. To test this hypothesis, we investigated: (i) the effect of HFE antisense oligonucleotides on apical iron uptake by polarized Caco-2 cells; (ii) the apical/basolateral membrane distribution of HFE, β-2 microglobulin and DMT1; (iii) the putative molecular association between HFE and DMT1. We found that HFE antisense treatment reduced HFE expression and increased apical iron uptake, whereas transfection with wild-type HFE inhibited iron uptake. Thus, an inverse relationship was established between HFE levels and apical iron uptake activity. Selective apical or basolateral biotinylation indicated preferential localization of DMT1 to the apical membrane and of HFE and β-2 microglobulin (β2m) to the basolateral membrane. Ectopic expression of HFE resulted in increased distribution of HFE–β2m to the apical membrane. The amount of HFE–β2m in the apical membrane inversely correlated with apical iron uptake rates. Immunoprecipitations of HFE or β2m with specific antibodies resulted in the co-precipitation of DMT1. These results sustain a model by which direct interaction between DMT1 and HFE–β2m in the apical membrane of Caco-2 cells result in down-regulation of apical iron uptake activity.

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References

  • S Abboud DJ Haile (2000) ArticleTitleA novel mammalian iron-regulated protein involved in intracellular iron metabolism J Biol Chem 275 19906–19912 Occurrence Handle10747949 Occurrence Handle1:CAS:528:DC%2BD3cXkslahtbk%3D Occurrence Handle10.1074/jbc.M000713200

    Article  PubMed  CAS  Google Scholar 

  • M Arredondo P Muñoz CV Mura MT Núñez (2001) ArticleTitleHFE inhibits apical iron uptake by intestinal epithelial (Caco-2) cells FASEB J 15 1276–1278 Occurrence Handle11344112 Occurrence Handle1:CAS:528:DC%2BD3MXlvVOksbw%3D

    PubMed  CAS  Google Scholar 

  • KR Bridle DM Frazer SJ Wilkins JL Dixon DM Purdie DH Crawford VN Subramaniam LW Powell GJ Anderson GA Ramm (2003) ArticleTitleDisrupted hepcidin regulation in HFE-associated haemochromatosis and the liver as a regulator of body iron homoeostasis Lancet 361 669–673 Occurrence Handle12606179 Occurrence Handle1:CAS:528:DC%2BD3sXhsVajt7Y%3D Occurrence Handle10.1016/S0140-6736(03)12602-5

    Article  PubMed  CAS  Google Scholar 

  • P Davies C Enns (2004) ArticleTitleExpression of the hereditary hemochromatosis protein HFE increases ferritin levels by inhibiting iron export in HT29 cells J Biol Chem 279 25085–25092 Occurrence Handle15044462 Occurrence Handle1:CAS:528:DC%2BD2cXksFyhur0%3D Occurrence Handle10.1074/jbc.M400537200

    Article  PubMed  CAS  Google Scholar 

  • RE Fleming MC Migas X Zhou J Jiang RS Britton ME Brunt S Tomatsu A Waheed BR Bacon WS Sly (1999) ArticleTitleMechanism of increased iron absorption in murine model of hereditary hemochromatosis: increased duodenal expression of the iron transporter DMT1 Proc Natl Acad Sci USA 96 3143–3148 Occurrence Handle10077651 Occurrence Handle1:CAS:528:DyaK1MXhvFykt7Y%3D Occurrence Handle10.1073/pnas.96.6.3143

    Article  PubMed  CAS  Google Scholar 

  • DM Frazer CD Vulpe AT McKie SJ Wilkins D Trinder GJ Cleghorn GJ Anderson (2001) ArticleTitleCloning and gastrointestinal expression of rat hephaestin: relationship to other iron transport proteins Am J Physiol 281 G931–G939 Occurrence Handle1:CAS:528:DC%2BD3MXnsFSksLc%3D

    CAS  Google Scholar 

  • DM Frazer SJ Wilkins EM Becker CD Vulpe AT McKie D Trinder GJ Anderson (2002) ArticleTitleHepcidin expression inversely correlates with the expression of duodenal iron transporters and iron absorption in rats Gastroenterologia 123 835–844 Occurrence Handle1:CAS:528:DC%2BD38Xntl2murs%3D Occurrence Handle10.1053/gast.2002.35353

    Article  CAS  Google Scholar 

  • DM Frazer GJ Anderson (2003) ArticleTitleThe orchestration of body iron intake: how and where do enterocytes receive their cues? Blood Cells Mol Dis 30 288–297 Occurrence Handle12737947 Occurrence Handle1:CAS:528:DC%2BD3sXjsVGmtrs%3D Occurrence Handle10.1016/S1079-9796(03)00039-1

    Article  PubMed  CAS  Google Scholar 

  • MA Gárate MT Núñez (2000) ArticleTitleOverexpression of the ferritin iron-responsive element decreases the labile iron pool and abolishes the regulation of iron absorption by intestinal epithelial (Caco-2) cells J Biol Chem 275 1651–1655 Occurrence Handle10636858 Occurrence Handle10.1074/jbc.275.3.1651

    Article  PubMed  Google Scholar 

  • MD Garrick KG Dolan C Horbinski AJ Ghio D Higgins M Porubcin EG Moore LN Hainsworth JN Umbreit ME Conrad L Feng A Lis JA Roth S Singleton LM Garrick (2003) ArticleTitleDMT1: A mammalian transporter for multiple metals BioMetals 16 41–54 Occurrence Handle12572663 Occurrence Handle1:CAS:528:DC%2BD38XnvFOhsrk%3D Occurrence Handle10.1023/A:1020702213099

    Article  PubMed  CAS  Google Scholar 

  • H Gunshin B Mackenzie UV Berger Y Gunshin MF Romero WF Boron S Nussberger JL Gollan MA Hediger (1997) ArticleTitleCloning and characterization of a mammalian proton-coupled metal-ion transporter Nature 388 482–488 Occurrence Handle9242408 Occurrence Handle1:CAS:528:DyaK2sXltFGqtbY%3D Occurrence Handle10.1038/41343

    Article  PubMed  CAS  Google Scholar 

  • A Le Bivic A Quaroni B Nichols E Rodriguez-Boulan (1990) ArticleTitleBiogenetic pathway of plasma membrane proteins in Caco-2 cells, a human intestinal epithelial cell line J Cell Biol 111 1351–1361 Occurrence Handle1976637 Occurrence Handle1:CAS:528:DyaK3cXls12ntLw%3D Occurrence Handle10.1083/jcb.111.4.1351

    Article  PubMed  CAS  Google Scholar 

  • JE Levy LK Montross NC Andrews (2000) ArticleTitleGenes that modify the hemochromatosis phenotype in mice J Clin Invest 105 1209–1216 Occurrence Handle10791995 Occurrence Handle1:CAS:528:DC%2BD3cXivFKgtr4%3D Occurrence Handle10.1172/JCI9635

    Article  PubMed  CAS  Google Scholar 

  • L Mahraoui M Ruosset E Dussaulx A Darmoul A Zweibaun E Brot-Laroche (1992) ArticleTitleExpression and localization of GLUT-5 in Caco-2 cells, human small intestine, and colon Am J Physiol 263 G312–G318 Occurrence Handle1384349 Occurrence Handle1:CAS:528:DyaK38XmtV2ntbw%3D

    PubMed  CAS  Google Scholar 

  • AT McKie D Barrow GO Latunde-Dada A Rolfs G Sager E Mudaly M Mudaly C Richardson D Barlow A Bomford TJ Peters KB Raja S Shirali MA Hediger F Farzaneh RJ Simpson (2001) ArticleTitleAn iron-regulated ferric reductase associated with the absorption of dietary iron Science 291 1755–1759 Occurrence Handle11230685 Occurrence Handle1:CAS:528:DC%2BD3MXhslSkurY%3D Occurrence Handle10.1126/science.1057206

    Article  PubMed  CAS  Google Scholar 

  • E Nemeth MS Tuttle J Powelson MB Vaughn A Donovan DM Ward T Ganz J Kaplan (2004) ArticleTitleHepcidin regulates cellular iron efflux by binding to ferroportin and inducing its internalization Science 306 2090–2093 Occurrence Handle15514116 Occurrence Handle1:CAS:528:DC%2BD2cXhtVOjsL3K Occurrence Handle10.1126/science.1104742

    Article  PubMed  CAS  Google Scholar 

  • G Nicolas M Bennoun I Devaux C Beaumont B Grandchamp A Kahn S Vaulont (2001) ArticleTitleLack of hepcidin gene expression and severe tissue iron overload in upstream stimulatory factor 2 (USF2) knockout mice Proc Natl Acad Sci USA 98 8780–8785 Occurrence Handle11447267 Occurrence Handle1:CAS:528:DC%2BD3MXls1Witbs%3D Occurrence Handle10.1073/pnas.151179498

    Article  PubMed  CAS  Google Scholar 

  • G Nicolas M Bennoun A Porteu S Mativet C Beaumont B Grandchamp M Sirito M Sawadogo A Kahn S Vaulont (2002) ArticleTitleSevere iron deficiency anemia in transgenic mice expressing liver hepcidin Proc Natl Acad Sci USA 99 4596–4601 Occurrence Handle11930010 Occurrence Handle1:CAS:528:DC%2BD38XivFShu7o%3D Occurrence Handle10.1073/pnas.072632499

    Article  PubMed  CAS  Google Scholar 

  • MD Peterson MS Mooseker (1993) ArticleTitleAn in vitro model for the analysis of intestinal brush-border assembly. I. Ultrastructural analysis of cell-contact induced brush border assembly in Caco-2 cells J Cell Sci 105 445–460 Occurrence Handle8408276

    PubMed  Google Scholar 

  • A Pietrangelo G Casalgrandi D Quaglino R Gualdi D Conte S Milani G Montosi L Cesarini E Ventura G Cairo (1995) ArticleTitleDuodenal ferritin synthesis in genetic hemochromatosis Gastroenterologia 108 208–217 Occurrence Handle1:STN:280:DyaK2M7gsVKgtA%3D%3D Occurrence Handle10.1016/0016-5085(95)90026-8

    Article  CAS  Google Scholar 

  • A Pietrangelo (2003) ArticleTitleHaemochromatosis Gut 52 IssueID(Suppl 2) ii23–ii30 Occurrence Handle12651879 Occurrence Handle1:CAS:528:DC%2BD3sXktVSksrk%3D

    PubMed  CAS  Google Scholar 

  • A Rolfs HL Bonkovsky JG Kohlroser K McNeal A Sharma UV Berger MA Hediger (2002) ArticleTitleIntestinal expression of gene involved in iron absorption in humans Am J Physiol 282 G598–G607 Occurrence Handle1:CAS:528:DC%2BD38XivVelsLg%3D

    CAS  Google Scholar 

  • CN Roy DM Penny JN Feder CA Enns (1999) ArticleTitleThe hereditary hemochromatosis protein, HFE, specifically regulates transferrin-mediated iron uptake in HeLa cells J Biol Chem 274 9022–9028 Occurrence Handle10085150 Occurrence Handle1:CAS:528:DyaK1MXitFyrurk%3D Occurrence Handle10.1074/jbc.274.13.9022

    Article  PubMed  CAS  Google Scholar 

  • V Tapia M Arredondo MT Núñez (1996) ArticleTitleRegulation of Fe absorption by cultured intestinal (Caco-2) cell monolayers with varied Fe status Am J Physiol 271 G443–G447 Occurrence Handle8843768 Occurrence Handle1:CAS:528:DyaK28XmtFajtbg%3D

    PubMed  CAS  Google Scholar 

  • A Waheed S Parkkila XY Zhou S Tomatsu Z Tsuchihashi JN Feder RC Schatzman RS Britton BR Bacon WS Sly (1997) ArticleTitleHereditary hemochromatosis: effects of C282Y and H63D mutations on association with β 2-microglobulin, intracellular processing, and cell surface expression of the HFE protein in COS-7 cells Proc Natl Acad Sci USA 94 12384–12389 Occurrence Handle9356458 Occurrence Handle1:CAS:528:DyaK2sXns1ylsLo%3D Occurrence Handle10.1073/pnas.94.23.12384

    Article  PubMed  CAS  Google Scholar 

  • J Wang G Chen K Pantopoulos (2002) ArticleTitleThe hemochromatosis protein HFE induces an apparent iron-deficient phenotype in H1299 cells that is not corrected by co-expression of beta2-microglobulin Biochem J 370 891–899 Occurrence Handle10.1042/BJ20021607 Occurrence Handle1:CAS:528:DC%2BD3sXhvFKhsrc%3D

    Article  CAS  Google Scholar 

  • S Yamaji P Sharp B Ramesh SK Srai (2004) ArticleTitleInhibition of iron transport across human intestinal epithelial cells by hepcidin Blood 104 2178–2178 Occurrence Handle15178582 Occurrence Handle1:CAS:528:DC%2BD2cXotVOqu7k%3D Occurrence Handle10.1182/blood-2004-03-0829

    Article  PubMed  CAS  Google Scholar 

  • KY Yeh M Yeh J Glass (2004) ArticleTitleHepcidin regulation of ferroportin 1 expression in the liver and intestine of the rat Am J Physiol 286 G385–G394 Occurrence Handle1:CAS:528:DC%2BD2cXitlSgt7o%3D Occurrence Handle10.1152/ajprenal.00226.2003

    Article  CAS  Google Scholar 

  • H Zoller RO Koch I Theurl P Obrist A Pietrangelo G Montosi DJ Haile W Vogel G Weiss (2001) ArticleTitleExpression of the duodenal iron transporters divalent-metal transporter 1 and Ireg 1 in iron deficiency and iron overload Gastroenterologia 120 1412–1419 Occurrence Handle1:CAS:528:DC%2BD3MXjs1Srt7o%3D Occurrence Handle10.1053/gast.2001.24033

    Article  CAS  Google Scholar 

  • C Zurzolo A Le Bivic E Rodriguez-Boulan (1994) Cell surface biotinylation techniques JE Celis (Eds) Cell Biology: A Laboratory Handbook NumberInSeries3 Academic Press San Diego, CA

    Google Scholar 

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Authors and Affiliations

  1. Micronutrients Laboratory, Institute of Nutrition and Food Technology (INTA), Universidad de Chile, Macul 5540, Macul, Santiago, Chile

    Miguel Arredondo

  2. Department of Biology, Faculty of Sciences, and Millennium Institute for Advanced Studies in Cell Biology and Biotechnology, Universidad de Chile, Macul 5540, Macul, Santiago, Chile

    Victoria Tapia, Alejandro Rojas, Pabla Aguirre, Francisca Reyes, Maria Paz Marzolo & Marco T. Núñez

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  1. Miguel Arredondo
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  2. Victoria Tapia
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  3. Alejandro Rojas
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  4. Pabla Aguirre
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  5. Francisca Reyes
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  6. Maria Paz Marzolo
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  7. Marco T. Núñez
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Correspondence to Miguel Arredondo.

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Arredondo, M., Tapia, V., Rojas, A. et al. Apical distribution of HFE–β2-microglobulin is associated with inhibition of apical iron uptake in intestinal epithelia cells. Biometals 19, 379–388 (2006). https://doi.org/10.1007/s10534-005-6687-x

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  • DOI: https://doi.org/10.1007/s10534-005-6687-x

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