Plant Molecular Biology

, Volume 40, Issue 1, pp 37–44 | Cite as

The IRT1 protein from Arabidopsis thaliana is a metal transporter with a broad substrate range

  • Yulia O. Korshunova
  • David Eide
  • W. Gregg Clark
  • Mary Lou Guerinot
  • Himadri B. Pakrasi


The molecular basis for the transport of manganese across membranes in plant cells is poorly understood. We have found that IRT1, an Arabidopsis thaliana metal ion transporter, can complement a mutant Saccharomyces cerevisiae strain defective in high-affinity manganese uptake (smf1Δ). The IRT1 protein has previously been identified as an iron transporter. The current studies demonstrated that IRT1, when expressed in yeast, can transport manganese as well. This manganese uptake activity was inhibited by cadmium, iron(II) and zinc, suggesting that IRT1 can transport these metals. The IRT1 cDNA also complements a zinc uptake-deficient yeast mutant strain (zrt1zrt2), and IRT1-dependent zinc transport in yeast cells is inhibited by cadmium, copper, cobalt and iron(III). However, IRT1 did not complement a copper uptake-deficient yeast mutant (ctr1), implying that this transporter is not involved in the uptake of copper in plant cells. The expression of IRT1 is enhanced in A. thaliana plants grown under iron deficiency. Under these conditions, there were increased levels of root-associated manganese, zinc and cobalt, suggesting that, in addition to iron, IRT1 mediates uptake of these metals into plant cells. Taken together, these data indicate that the IRT1 protein is a broad-range metal ion transporter in plants.

copper iron manganese metal transport plant root zinc 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Andersson B, Styring S: Photosystem II: molecular organization, function and acclimation. Curr Top Bioenerg 16: 1–81 (1991).Google Scholar
  2. 2.
    Anderson JA, Huprikar SS, Kochian LV, Lucas WJ, Gaber RF: Functional expression of a probable Arabidopsis thaliana potassium channel in Saccharomyces cerevisiae. Proc Natl Acad Sci USA 89: 3736–3740 (1992).Google Scholar
  3. 3.
    Bartsevich VV, Pakrasi HB: Molecular identification of an ABC transporter complex for manganese: analysis of a cyanobacterial mutant strain impaired in the photosynthetic oxygen evolution process. EMBO J 14: 1845–1853 (1995).Google Scholar
  4. 4.
    Bartsevich VV, Pakrasi HB: Manganese transport in the cyanobacterium Synechocystis sp. PCC 6803. J Biol Chem 271: 26057–26061 (1996).Google Scholar
  5. 5.
    Belouchi AM, Cellier M, Kwan T, Saini B, Leroux G, Gros P: The macrophage specific membrane protein Nramp controlling natural resistance to infections in mice has homologues expressed in the root system of plants. Plant Mol Biol 29: 1181–1196 (1995).Google Scholar
  6. 6.
    Belouchi AM, Kwan T, Gros P: Cloning and characterization of the OsNramp family from Oryza sativa, a new family of membrane proteins possibly implicated in the transport of metal ions. Plant Mol Biol 33: 1085–1092 (1997).Google Scholar
  7. 7.
    Boeke JD, Trueheart J, Natsoulis G, Fink GR: 5-Fluoroorotic acid as a selective agent in yeast molecular genetics. Meth Enzymol 154: 164–175 (1987).Google Scholar
  8. 8.
    Cohen CK, Fox TC, Garvin DF, Kochian LV: The role of iron-deficiency stress responses in stimulating heavy-metal transport in plants. Plant Physiol 116: 1063–1072 (1998).Google Scholar
  9. 9.
    Culotta Cizeweski V, Klomp LWJ, Strain J, Casareno RLB, Krems B, Gitlin JD: The copper chaperone for superoxide dismutase. J Biol Chem 272: 23469–23472 (1997).Google Scholar
  10. 10.
    Dancis A, Yuan DS, Haile D, Askwith C, Eide D, Moehle C, Kaplan J, Klausner DR: Molecular characterization of a copper transport protein in S. cerevisiae: an unexpected role for copper in iron transport. Cell 76: 393–402 (1994).Google Scholar
  11. 11.
    Eide D, Broderius M, Fett J, Guerinot ML: A novel ironregulated metal transporter from plants identified by functional expression in yeast. Proc Natl Acad Sci USA 93: 5624–5628 (1996).Google Scholar
  12. 12.
    Feinberg AP, Vogelstein B: A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 137: 266–267 (1984).Google Scholar
  13. 13.
    Frausto da Silva JJR, Williams RJP: The Biological Chemistry of the Elements: The Inorganic Chemistry of Life. Oxford University Press, New York (1991).Google Scholar
  14. 14.
    Glerum DM, Shtanko A, Tzagoloff A: Characterization of COX17, a yeast gene involved in copper metabolism and assembly of cytochrome oxidase. J Biol Chem 271: 14504–14509 (1996).Google Scholar
  15. 15.
    Grotz N, Fox T, Connolly E, Park W, Guerinot ML, Eide D: Identification of a family of zinc transporter genes from Arabidopsis that respond to zinc deficiency. Proc Natl Acad Sci USA 95: 7220–7224 (1998).Google Scholar
  16. 16.
    Gunshin H, Mackenzie B, Berger UV, Gunshin Y, Romero MF, Boron WF, Nussberger S, Gollan JL, Hedlger MA: Cloning and characterization of a mammalian proton-coupled metalion transporter. Nature 388: 482–484 (1997).Google Scholar
  17. 17.
    Kampfenkel K, Kushnir S, Babiychuk E, Inze D, Van Montagu MV:Molecular characterization of a putative Arabidopsis thaliana copper transporter and its yeast homologue. J Biol Chem 270: 28479–28486 (1995).Google Scholar
  18. 18.
    Kaufman R, Swaroop M, Murtha-Riel P: Depletion of manganese within the secretory pathway inhibits o-linked glycosylation in mammalian cells. Biochemistry 33: 9813–9819 (1994).Google Scholar
  19. 19.
    Larson EJ, Pecoraro VL: Introduction to manganese enzymes. In: Pecoraro VL <nt>(ed.)</nt>, Manganese Redox Enzymes. VCH Publishers, New York, pp. 1–28 (1992).Google Scholar
  20. 20.
    Lin S-J, Culotta Cizeweski V: The ATX1 gene of Saccharomyces cerevisiae encodes a small metal homeostasis factor that protects cells against reactive oxygen toxicity. Proc Natl Acad Sci USA 92: 3784–3788 (1995).Google Scholar
  21. 21.
    Liu XF, Supek F, Nelson N, Culotta Cizewski V: Negative control of heavy metal uptake by the Saccharomyces cerevisiae BSD2 gene. J Biol Chem 272: 11763–11769 (1997).Google Scholar
  22. 22.
    Marschner H: Mineral Nutrition of Higher Plants. Academic Press, London (1986).Google Scholar
  23. 23.
    Minet M, Dufour M-E, Lacroute F: Complementation of Saccharomyces cerevisiae auxotrophic mutants by Arabidopsis thaliana cDNAs. Plant J 2: 417–422 (1992).Google Scholar
  24. 24.
    Murashige T, Skoog F: A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497 (1962).Google Scholar
  25. 25.
    Pilgrim ML, McClung R: Differential involvement of the circadian clock in the expression of genes required for ribulose-1,5-biphosphate carboxylase/oxygenase synthesis, assembly, and activation in Arabidopsis thaliana. Plant Physiol 103: 553–564 (1993).Google Scholar
  26. 26.
    Pufahl RA, Singer CP, Peariso KL, Lin S-J, Schmidt PJ, Fahrni CJ, Culotta Cizeweski V, Hahn-Penner JE, O'Halloran TV: Metal ion chaperone function of the soluble Cu(I) receptor Atx1. Science 278: 853–855 (1997).Google Scholar
  27. 27.
    Rodecap KD, Tingey DT, Lee EH: Iron nutrition influence on cadmium accumulation by Arabidopsis thaliana (L.) Heynh. J Environ Qual 23: 239–246 (1994).Google Scholar
  28. 28.
    Sambrook J, Fritsch E, Maniatis T: Molecular Cloning: A LaboratoryManual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).Google Scholar
  29. 29.
    Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).Google Scholar
  30. 30.
    Schachtman DP, Schroeder JI: Structure and transport mechanism of a high-affinity potassium uptake transporter from higher plants. Nature 370: 655–658 (1994).Google Scholar
  31. 31.
    Schiestl RH, Geitz RD: High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier. Curr Genet 16: 339–346 (1989).Google Scholar
  32. 32.
    Sentenac H, Bonneaud N, Minet M, Lacroute F, Salmon J-M, Gaymard F, Grignon C: Cloning and expression in yeast of a plant K+ transport system. Science 256: 663–665 (1992).Google Scholar
  33. 33.
    Sherman F, Fink GR, Hicks JB: Laboratory course manual for methods in yeast genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1986).Google Scholar
  34. 34.
    Strathern JN, Higgins DR: Recovery of plasmids from yeast into Escherichia coli shuttle vectors. Meth Enzymol 194: 319–328 (1991).Google Scholar
  35. 35.
    Supek F, Supekova L, Nelson H, Nelson N: Ayeast manganese transporter related to the macrophage protein involved in conferring resistance to mycobacteria. Proc Natl Acad Sci USA 93: 5105–5110 (1996).Google Scholar
  36. 36.
    Verwoerd TC, Dekker BMM, Hoekema A: A small-scale procedure for the rapid isolation of plant RNAs. Nucl Acids Res 17: 2362 (1989).Google Scholar
  37. 37.
    Welch RM, Norvell WA, Schaefer SC, Shaff JE, Kochian LV: Induction of iron(III) and copper(II) reduction in pea (Pisum sativum) roots by Fe and Cu status: does the root-cell plasmalemma Fe(III)-chelate reductase perform a general role in regulating cation uptake? Planta 190: 555–561 (1993).Google Scholar
  38. 38.
    Yi Y, Guerinot ML: Genetic evidence that induction of root Fe(III) chelate reductase activity is necessary for iron uptake under iron deficiency. Plant J 10: 835–844 (1996).Google Scholar
  39. 39.
    Zhao H, Eide D: The ZRT2 gene encodes the low affinity zinc transporter in Saccharomyces cerevisiae. J Biol Chem 271: 23203–23210 (1996).Google Scholar
  40. 40.
    Zhao H, Eide D: The yeast ZRT1 gene encodes the zinc transporter protein of high-affinity uptake system induced by zinc limitation. Proc Natl Acad Sci USA 93: 2454–2458 (1996).Google Scholar

Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • Yulia O. Korshunova
    • 1
  • David Eide
    • 2
  • W. Gregg Clark
    • 1
  • Mary Lou Guerinot
    • 3
  • Himadri B. Pakrasi
    • 1
  1. 1.Department of BiologyWashington UniversitySt. LouisUSA
  2. 2.Nutritional Science ProgramUniversity of MissouriColumbiaUSA
  3. 3.Department of Biological SciencesDartmouth CollegeHanoverUSA

Personalised recommendations