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. 1996 Jul 1;15(13):3377–3384.

Iron-regulated DNA binding by the AFT1 protein controls the iron regulon in yeast.

Y Yamaguchi-Iwai 1, R Stearman 1, A Dancis 1, R D Klausner 1
PMCID: PMC451901  PMID: 8670839

Abstract

Iron deprivation of Saccharomyces cerevisiae induces transcription of genes required for high-affinity iron uptake. AFT1 mediates this transcriptional control. In this report, the 5'-flanking region of FET3, which encodes a copper-dependent oxidase required for iron transport, was analyzed and found to contain a DNA sequence responsible for AFT1-regulated gene expression. AFT1 was capable of interacting specifically with this DNA sequence. A core element within this DNA sequence necessary for the binding of AFT1 was also determined. In vivo footprinting demonstrated occupancy of the AFT1 binding site in cells deprived of iron and not in cells grown in the presence of iron. Thus, the environmental signal resulting from iron deprivation was transduced through the regulated binding of AFT1 to the FET3 promoter, followed by the activation of transcription. A regulon of genes under the control of AFT1 could be defined. AFT1 was able to bind to a consensus binding site (PyPuCACCCPu) in the 5' region of FRE1, FRE2, FTR1, FTH1 and CCC2.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Askwith C., Eide D., Van Ho A., Bernard P. S., Li L., Davis-Kaplan S., Sipe D. M., Kaplan J. The FET3 gene of S. cerevisiae encodes a multicopper oxidase required for ferrous iron uptake. Cell. 1994 Jan 28;76(2):403–410. doi: 10.1016/0092-8674(94)90346-8. [DOI] [PubMed] [Google Scholar]
  2. Bagg A., Neilands J. B. Molecular mechanism of regulation of siderophore-mediated iron assimilation. Microbiol Rev. 1987 Dec;51(4):509–518. doi: 10.1128/mr.51.4.509-518.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dancis A., Roman D. G., Anderson G. J., Hinnebusch A. G., Klausner R. D. Ferric reductase of Saccharomyces cerevisiae: molecular characterization, role in iron uptake, and transcriptional control by iron. Proc Natl Acad Sci U S A. 1992 May 1;89(9):3869–3873. doi: 10.1073/pnas.89.9.3869. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dancis A., Yuan D. S., Haile D., Askwith C., Eide D., Moehle C., Kaplan J., Klausner R. D. Molecular characterization of a copper transport protein in S. cerevisiae: an unexpected role for copper in iron transport. Cell. 1994 Jan 28;76(2):393–402. doi: 10.1016/0092-8674(94)90345-x. [DOI] [PubMed] [Google Scholar]
  5. De Silva D. M., Askwith C. C., Eide D., Kaplan J. The FET3 gene product required for high affinity iron transport in yeast is a cell surface ferroxidase. J Biol Chem. 1995 Jan 20;270(3):1098–1101. doi: 10.1074/jbc.270.3.1098. [DOI] [PubMed] [Google Scholar]
  6. Dumitru I., McNeil J. B. A simple in vivo footprinting method to examine DNA-protein interactions over the yeast PYK UAS element. Nucleic Acids Res. 1994 Apr 25;22(8):1450–1455. doi: 10.1093/nar/22.8.1450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eide D., Davis-Kaplan S., Jordan I., Sipe D., Kaplan J. Regulation of iron uptake in Saccharomyces cerevisiae. The ferrireductase and Fe(II) transporter are regulated independently. J Biol Chem. 1992 Oct 15;267(29):20774–20781. [PubMed] [Google Scholar]
  8. Fu D., Beeler T. J., Dunn T. M. Sequence, mapping and disruption of CCC2, a gene that cross-complements the Ca(2+)-sensitive phenotype of csg1 mutants and encodes a P-type ATPase belonging to the Cu(2+)-ATPase subfamily. Yeast. 1995 Mar;11(3):283–292. doi: 10.1002/yea.320110310. [DOI] [PubMed] [Google Scholar]
  9. Georgatsou E., Alexandraki D. Two distinctly regulated genes are required for ferric reduction, the first step of iron uptake in Saccharomyces cerevisiae. Mol Cell Biol. 1994 May;14(5):3065–3073. doi: 10.1128/mcb.14.5.3065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Guarente L., Ptashne M. Fusion of Escherichia coli lacZ to the cytochrome c gene of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2199–2203. doi: 10.1073/pnas.78.4.2199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Guarente L. Yeast promoters and lacZ fusions designed to study expression of cloned genes in yeast. Methods Enzymol. 1983;101:181–191. doi: 10.1016/0076-6879(83)01013-7. [DOI] [PubMed] [Google Scholar]
  12. Guo B., Yu Y., Leibold E. A. Iron regulates cytoplasmic levels of a novel iron-responsive element-binding protein without aconitase activity. J Biol Chem. 1994 Sep 30;269(39):24252–24260. [PubMed] [Google Scholar]
  13. Haile D. J., Rouault T. A., Harford J. B., Kennedy M. C., Blondin G. A., Beinert H., Klausner R. D. Cellular regulation of the iron-responsive element binding protein: disassembly of the cubane iron-sulfur cluster results in high-affinity RNA binding. Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):11735–11739. doi: 10.1073/pnas.89.24.11735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jungmann J., Reins H. A., Lee J., Romeo A., Hassett R., Kosman D., Jentsch S. MAC1, a nuclear regulatory protein related to Cu-dependent transcription factors is involved in Cu/Fe utilization and stress resistance in yeast. EMBO J. 1993 Dec 15;12(13):5051–5056. doi: 10.1002/j.1460-2075.1993.tb06198.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Karlin K. D. Metalloenzymes, structural motifs, and inorganic models. Science. 1993 Aug 6;261(5122):701–708. doi: 10.1126/science.7688141. [DOI] [PubMed] [Google Scholar]
  16. Klausner R. D., Rouault T. A., Harford J. B. Regulating the fate of mRNA: the control of cellular iron metabolism. Cell. 1993 Jan 15;72(1):19–28. doi: 10.1016/0092-8674(93)90046-s. [DOI] [PubMed] [Google Scholar]
  17. Köhrer K., Domdey H. Preparation of high molecular weight RNA. Methods Enzymol. 1991;194:398–405. doi: 10.1016/0076-6879(91)94030-g. [DOI] [PubMed] [Google Scholar]
  18. Mitchell P. J., Tjian R. Transcriptional regulation in mammalian cells by sequence-specific DNA binding proteins. Science. 1989 Jul 28;245(4916):371–378. doi: 10.1126/science.2667136. [DOI] [PubMed] [Google Scholar]
  19. O'Halloran T. V. Transition metals in control of gene expression. Science. 1993 Aug 6;261(5122):715–725. doi: 10.1126/science.8342038. [DOI] [PubMed] [Google Scholar]
  20. Samaniego F., Chin J., Iwai K., Rouault T. A., Klausner R. D. Molecular characterization of a second iron-responsive element binding protein, iron regulatory protein 2. Structure, function, and post-translational regulation. J Biol Chem. 1994 Dec 9;269(49):30904–30910. [PubMed] [Google Scholar]
  21. Stearman R., Yuan D. S., Yamaguchi-Iwai Y., Klausner R. D., Dancis A. A permease-oxidase complex involved in high-affinity iron uptake in yeast. Science. 1996 Mar 15;271(5255):1552–1557. doi: 10.1126/science.271.5255.1552. [DOI] [PubMed] [Google Scholar]
  22. Verma I. M., Stevenson J. K., Schwarz E. M., Van Antwerp D., Miyamoto S. Rel/NF-kappa B/I kappa B family: intimate tales of association and dissociation. Genes Dev. 1995 Nov 15;9(22):2723–2735. doi: 10.1101/gad.9.22.2723. [DOI] [PubMed] [Google Scholar]
  23. Voisard C., Wang J., McEvoy J. L., Xu P., Leong S. A. urbs1, a gene regulating siderophore biosynthesis in Ustilago maydis, encodes a protein similar to the erythroid transcription factor GATA-1. Mol Cell Biol. 1993 Nov;13(11):7091–7100. doi: 10.1128/mcb.13.11.7091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Yamaguchi-Iwai Y., Dancis A., Klausner R. D. AFT1: a mediator of iron regulated transcriptional control in Saccharomyces cerevisiae. EMBO J. 1995 Mar 15;14(6):1231–1239. doi: 10.1002/j.1460-2075.1995.tb07106.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Yuan D. S., Stearman R., Dancis A., Dunn T., Beeler T., Klausner R. D. The Menkes/Wilson disease gene homologue in yeast provides copper to a ceruloplasmin-like oxidase required for iron uptake. Proc Natl Acad Sci U S A. 1995 Mar 28;92(7):2632–2636. doi: 10.1073/pnas.92.7.2632. [DOI] [PMC free article] [PubMed] [Google Scholar]

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