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Evaluation of the iron regulatory protein-1 interactome

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Abstract

The interactions of iron regulatory proteins (IRPs) with mRNAs containing an iron-responsive element (IRE) is a major means through which intracellular iron homeostasis is maintained and integrated with cellular function. Although IRE–IRP interactions have been proposed to modulate the expression of a diverse number of mRNAs, a transcriptome analysis of the interactions that form within the native mRNA structure and cellular environment has not previously been described. An RNA-CLIP study is described here that identified IRP-1 interactions occurring within a primary cell line expressing physiologically relevant amounts of mRNA and protein. The study suggests that only a small subset of the previously proposed IREs interact with IRP-1 in situ. Identifying authentic IRP interactions is not only important to a greater understanding of iron homeostasis and its integration with cell biology but also to the development of novel therapeutics that can compensate for iron imbalances.

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References

  • Abboud S, Haile DJ (2000) A novel mammalian iron-regulated protein involved in intracellular iron metabolism. J Biol Chem 275:19906–19912

    Article  CAS  PubMed  Google Scholar 

  • Ande SR, Mishra S (2011) Nuclear coded mitochondrial protein prohibitin is an iron regulated iron binding protein. Mitochondrion 11:40–47

    Article  CAS  PubMed  Google Scholar 

  • Basilion JP, Rouault TA, Massinople CM, Klausner RD, Burgess WH (1994) The iron-responsive element-binding protein: localization of the RNA-binding site to the aconitase active-site cleft. Proc Natl Acad Sci 91:574–578

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Blankenberg D, Von Kuster G, Coraor N, Ananda G, Lazarus R, Mangan M, Nekrutenko A, Taylor J (2010) Galaxy: a web-based genome analysis tool for experimentalists. Curr Protoc Mol Biol 89:19.10.1–19.10.21

    Google Scholar 

  • Butt J, Kim HY, Basilion JP, Cohen S, Iwai K, Philpott CC, Altschul S, Klausner RD, Rouault TA (1996) Differences in the RNA binding sites of iron regulatory proteins and potential target diversity. Proc Natl Acad Sci 93:4345–4349

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Campillos M, Cases I, Hentze MW, Sanchez M (2010) SIREs: searching for iron-responsive elements. Nucleic Acids Res 38:W360–W367

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Casey JL, Hentze MW, Koeller DM, Caughman SW, Rouault TA, Klausner RD, Harford JB (1988) Iron-responsive elements: regulatory RNA sequences that control mRNA levels and translation. Science 240:924–928

    Article  CAS  PubMed  Google Scholar 

  • Cho H-H, Cahill CM, Vanderburg CR, Scherzer CR, Wang B, Huang X, Rogers JT (2010) Selective translational control of the Alzheimer amyloid precursor protein transcript by iron regulatory protein-1. J Biol Chem 285:31217–31232

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cmejla R, Petrak J, Cmejlova J (2006) A novel iron responsive element in the 3′UTR of human MRCKalpha. Biochem Biophys Res Commun 341:158–166

    Article  CAS  PubMed  Google Scholar 

  • Crielaard BJ, Lammers T, Rivella S (2017) Targeting iron metabolism in drug discovery and delivery. Nat Rev Drug Discov 16:400–423

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dandekar T, Stripecke R, Gray NK, Goossen B, Constable A, Johansson HE, Hentze MW (1991) Identification of a novel iron-responsive element in murine and human erythroid delta-aminolevulinic acid synthase mRNA. EMBO J 10:1903–1909

    CAS  PubMed  PubMed Central  Google Scholar 

  • Dix DJ, Lin PN, McKenzie AR, Walden WE, Theil EC (1993) The influence of the base-paired flanking region on structure and function of the ferritin mRNA iron regulatory element. J Mol Biol 231:230–240

    Article  CAS  PubMed  Google Scholar 

  • Donovan A, Brownlie A, Zhou Y, Shepard J, Pratt SJ, Moynihan J, Paw BH, Drejer A, Barut B, Zapata A et al (2000) Positional cloning of zebrafish ferroportin1 identifies a conserved vertebrate iron exporter. Nature 403:776–781

    Article  CAS  PubMed  Google Scholar 

  • dos Santos CO, Dore LC, Valentine E, Shelat SG, Hardison RC, Ghosh M, Wang W, Eisenstein RS, Costa FF, Weiss MJ (2008) An iron responsive element-like stem-loop regulates alpha-hemoglobin-stabilizing protein mRNA. J Biol Chem 283:26956–26964

    Article  PubMed  PubMed Central  Google Scholar 

  • Giardine B, Riemer C, Hardison RC, Burhans R, Elnitski L, Shah P, Zhang Y, Blankenberg D, Albert I, Taylor J et al (2005) Galaxy: a platform for interactive large-scale genome analysis. Genome Res 15:1451–1455

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Goecks J, Nekrutenko A, Taylor J, Galaxy T (2010) Galaxy: a comprehensive approach for supporting accessible, reproducible, and transparent computational research in the life sciences. Genome Biol 11:R86

    Article  PubMed  PubMed Central  Google Scholar 

  • Gunshin H, Allerson CR, Polycarpou-Schwarz M, Rofts A, Rogers JT, Kishi F, Hentze W, Rouault TA, Andrews NC, Hediger MA (2001) Iron-dependent regulation of the divalent metal ion transporter. FEBS Lett 509:309–316

    Article  CAS  PubMed  Google Scholar 

  • Guo JU, Bartel DP (2016) RNA G-quadruplexes are globally unfolded in eukaryotic cells and depleted in bacteria. Science 353:23

    Article  Google Scholar 

  • Henderson BR, Menotti E, Bonnard C, Kuhn LC (1994) Optimal sequence and structure of iron-responsive elements. Selection of RNA stem-loops with high affinity for iron regulatory factor. J Biol Chem 269:17481–17489

    CAS  PubMed  Google Scholar 

  • Henderson BR, Menotti E, Kuhn LC (1996) Iron regulatory proteins 1 and 2 bind distinct sets of RNA target sequences. J Biol Chem 271:4900–4908

    Article  CAS  PubMed  Google Scholar 

  • Hentze MW, Caughman SW, Rouault TA, Barriocanal JG, Dancis A, Harford JB, Klausner RD (1987) Identification of the iron-responsive element for the translational regulation of human ferritin mRNA. Science 238:1570–1573

    Article  CAS  PubMed  Google Scholar 

  • Hu Z, Gulec S, Collins JF (2010) Cross-species comparison of genomewide gene expression profiles reveals induction of hypoxia-inducible factor-responsive genes in iron-deprived intestinal epithelial cells. Am J Physiol Cell Physiol 299:C930–C938

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Khan MA, Walden WE, Theil EC, Goss DJ (2017) Thermodynamic and kinetic analyses of iron response element (IRE)-mRNA binding to iron regulatory protein, IRP1. Scientific Reports 7:8532

    Article  PubMed  PubMed Central  Google Scholar 

  • Kohler SA, Menotti E, Kuhn LC (1999) Molecular cloning of mouse glycolate oxidase. High evolutionary conservation and presence of an iron-responsive element-like sequence in the mRNA. J Biol Chem 274:2401–2407

    Article  CAS  PubMed  Google Scholar 

  • Leibold EA, Laudano A, Yu Y (1990) Structural requirements of iron-responsive elements for binding of the protein involved in both transferrin receptor and ferritin mRNA post-transcriptional regulation. Nucleic Acids Res 18:1819–1824

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lin E, Graziano JH, Freyer GA (2001) Regulation of the 75-kDa subunit of mitochondrial complex I by iron. J Biol Chem 276:27685–27692

    Article  CAS  PubMed  Google Scholar 

  • Liu Z, Lanford R, Mueller S, Gerhard GS, Luscieti S, Sanchez M, Devireddy L (2012) Siderophore-mediated iron trafficking in humans is regulated by iron. J Mol Med 90:1209–1221

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Luscieti S, Galy B, Gutierrez L, Reinke M, Couso J, Shvartsman M, Di Pascale A, Witke W, Hentze MW, Pilo Boyl P et al (2017) The actin binding protein profilin 2 is a novel regulator of iron homeostasis. Blood 130:1934–1945

    Article  PubMed  Google Scholar 

  • McKie AT, Marciani P, Rolfs A, Brennan K, Wehr K, Barrow D, Miret S, Bomford A, Peters TJ, Farzaneh F et al (2000) A novel duodenal iron-regulated transporter, IREG1, implicated in the basolateral transfer of iron to the circulation. Mol Cell 5:299–309

    Article  CAS  PubMed  Google Scholar 

  • Meehan HA, Connell GJ (2001) The hairpin loop but not the bulged C of the iron responsive element is essential for high affinity binding to iron regulatory protein-1. J Biol Chem 276:14791–14796

    Article  CAS  PubMed  Google Scholar 

  • Meyron-Holtz EG, Ghosh MC, Rouault TA (2004) Mammalian tissue oxygen levels modulate iron-regulatory protein activities in vivo. Science 306:2087–2090

    Article  CAS  PubMed  Google Scholar 

  • Mullner EW, Kuhn LC (1988) A stem-loop in the 3′untranslated region mediates iron-dependent regulation of transferrin receptor mRNA stability in the cytoplasm. Cell 53:815–825

    Article  CAS  PubMed  Google Scholar 

  • Papanikolaou G, Pantopoulos K (2017) Systemic iron homeostasis and erythropoiesis. IUBMB Life 69:399–413

    Article  CAS  PubMed  Google Scholar 

  • Rouskin S, Zubradt M, Washietl S, Kellis M, Weissman JS (2014) Genome-wide probing of RNA structure reveals active unfolding of mRNA structures in vivo. Nature 505:701–705

    Article  CAS  PubMed  Google Scholar 

  • Rupani DN, Connell GJ (2016) Transferrin receptor mRNA interactions contributing to iron homeostasis. RNA 22:1271–1282

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sanchez M, Galy B, Dandekar T, Bengert P, Vainshtein Y, Stolte J, Muckenthaler MU, Hentze MW (2006) Iron regulation and the cell cycle: identification of an iron-responsive element in the 3′-untranslated region of human cell division cycle 14A mRNA by a refined microarray-based screening strategy. J Biol Chem 281:22865–22874

    Article  CAS  PubMed  Google Scholar 

  • Sanchez M, Galy B, Muckenthaler MU, Hentze MW (2007) Iron-regulatory proteins limit hypoxia-inducible factor-2 alpha expression in iron deficiency. Nat Struct Mol 14:420–426

    Article  CAS  Google Scholar 

  • Sanchez M, Galy B, Schwanhaeusser B, Blake J, Bahr-Ivacevic T, Benes V, Selbach M, Muckenthaler MU, Hentze MW (2011) Iron regulatory protein-1 and -2: transcriptome-wide definition of binding mRNAs and shaping of the cellular proteome by iron regulatory proteins. Blood 118:e168–e179

    Article  CAS  PubMed  Google Scholar 

  • Schlegl J, Gegout V, Schlager B, Hentze MW, Westhof E, Ehresmann C, Ehresmann B, Romby P (1997) Probing the structure of the regulatory region of human transferrin receptor messenger RNA and its interaction with iron regulatory protein-1. RNA 3:1159–1172

    CAS  PubMed  PubMed Central  Google Scholar 

  • Spitale RC, Flynn RA, Zhang QC, Crisalli P, Lee B, Jung J-W, Kuchelmeister HY, Batista PJ, Torre EA, Kool ET et al (2015) Structural imprints in vivo decode RNA regulatory mechanisms. Nature 519:486–490

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ule J, Jensen KB, Ruggiu M, Mele A, Ule A, Darnell RB (2003) CLIP identifies Nova-regulated RNA networks in the brain. Science 302:1212–1215

    Article  CAS  PubMed  Google Scholar 

  • Walden WE, Selezneva AI, Dupuy J, Volbeda A, Fontecilla-Camps JC, Theil EC, Volz K (2006) Structure of dual function iron regulatory protein 1 complexed with ferritin IRE-RNA. Science 314:1903–1908

    Article  CAS  PubMed  Google Scholar 

  • Walden WE, Selezneva A, Volz K (2012) Accommodating variety in iron-responsive elements: crystal structure of transferrin receptor 1 B IRE bound to iron regulatory protein 1. FEBS Lett 586:32–35

    Article  CAS  PubMed  Google Scholar 

  • Zimmer M, Ebert BL, Neil C, Brenner K, Papaioannou I, Melas A, Tolliday N, Lamb J, Pantopoulos K, Golub T et al (2008) Small-molecule inhibitors of HIF-2a translation link its 5′UTR iron-responsive element to oxygen sensing. Mol Cell 32:838–848

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was funded in part by a grant from the University of Minnesota Academic Health Center to GJC (Grant Number 12.04). We also acknowledge the assistance of the University of Minnesota Genomics Center and the University of Minnesota Supercomputing Institute.

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This work was supported in part by a grant from the University of Minnesota Academic Health Center to GJC (Grant Number 12.04).

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Correspondence to Gregory J. Connell.

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Connell, G.J., Danial, J.S. & Haastruthers, C.X. Evaluation of the iron regulatory protein-1 interactome. Biometals 31, 139–146 (2018). https://doi.org/10.1007/s10534-018-0076-8

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