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BioMetals

, Volume 24, Issue 1, pp 51–58 | Cite as

Characterization of zinc transport by divalent metal transporters of the ZIP family from the model legume Medicago truncatula

  • Brian W. Stephens
  • Douglas R. Cook
  • Michael A. GrusakEmail author
Article

Abstract

To understand how plants from the Fabaceae family maintain zinc (Zn) homeostasis, we have characterized the kinetics of three Zn transporting proteins from the ZIP family of divalent metal transporters in the model legume Medicago truncatula. Of six ZIP’s studied, MtZIP1, MtZIP5 and MtZIP6 were the only members from this family determined to transport Zn and were further characterized. MtZIP1 has a low affinity for Zn with a Km of 1 μM as compared to MtZIP5 and MtZIP6 that have a higher affinity for Zn with Km of 0.4 μM and 0.3 μM, respectively. Zn transport by MtZIP1 was more sensitive to inhibition by copper (Cu) concentrations than MtZIP5 and MtZIP6, because 3 μM Cu inhibited Zn transport by 80% in MtZIP1 while 5 μM Cu was required to achieve the same inhibition of Zn transport in MtZIP5 and MtZIP6. Cadmium (Cd) had a greater effect on the ability of MtZIP1 to transport Zn than MtZIP5 and MtZIP6, because at a concentration of 3 μM Cd, the Zn transport by MtZIP1 was inhibited 55% and the transport of Zn by MtZIP5 and MtZIP6 was inhibited by 20–30%. However, only MtZIP6 transported Cd at higher rates than those observed in the control plasmid pFL61, demonstrating a low affinity for Cd based on a Km of 57 μM. These results suggest that Medicago truncatula has both high and low affinity Zn transporters to maintain Zn homeostasis and that these transporters may function in different compartments within the plant.

Keywords

Zinc Membrane transport Kinetics Cadmium Copper Medicago truncatula 

Notes

Acknowledgments

This research was supported in part by HarvestPlus under Agreement number 58-6250-4-F029 and by the USDA-ARS under Agreement number 58-6250-6-003 to MAG. The contents of this publication do not necessarily reflect the views or policies of the US Department of Agriculture, nor does mention of trade names, commercial products, or organizations imply endorsement by the US Government.

References

  1. Bal W, Kozlowski H, Kupryszewski G, Mackiewicz Z, Pettit L, Robbins R (1993) Complexes of Cu(II) with Asn-Ser-Phe-Arg-Tyr-NH2; an example of metal ion-promoted conformational organization which results in exceptionally high complex stability. J Inorg Biochem 52:79–87CrossRefPubMedGoogle Scholar
  2. Berg JM, Shi Y (1996) The galvanization of biology: a growing appreciation for the roles of zinc. Science 271:1081–1085CrossRefPubMedGoogle Scholar
  3. Blaudez D, Kohler A, Martin F, Sanders D, Chalot M (2003) Poplar metal tolerance protein 1 confers zinc tolerance and is an oligomeric vacuolar zinc transporter with an essential leucine zipper motif. Plant Cell 15:2911–2928CrossRefPubMedGoogle Scholar
  4. Bowen JE (1969) Absorption of copper, zinc, and manganese by sugarcane leaf tissue. Plant Physiol 44:255–261CrossRefPubMedGoogle Scholar
  5. Broadley MR, White PJ, Hammond JP, Zelko I, Lux A (2007) Zinc in plants. New Phytol 173:677–702CrossRefPubMedGoogle Scholar
  6. Brown SL, Chaney RL, Angle JS, Baker AJM (1995) Zinc and cadmium uptake by hyperaccumulator thlaspi-caerulescens and metal-tolerant silene-vulgaris grown on sludge-amended soils. Environ Sci Technol 29:1581–1585CrossRefGoogle Scholar
  7. Burleigh SH, Kristensen BK, Bechmann IE (2003) A plasma membrane zinc transporter from Medicago truncatula is up-regulated in roots by Zn fertilization, yet down-regulated by arbuscular mycorrhizal colonization. Plant Mol Biol 52:1077–1088CrossRefPubMedGoogle Scholar
  8. Carroll M, Loneragan J (1968) Response of plant species to concentrations of zinc in solution. I. Growth and zinc content of plants. Aust J Agric Res 19:859–868CrossRefGoogle Scholar
  9. Cellier M, Prive G, Belouchi A, Kwan T, Rodrigues V, Chia W, Gros P (1995) Nramp defines a family of membrane proteins. Proc Natl Acad Sci USA 92:10089–10093CrossRefPubMedGoogle Scholar
  10. Chaudhry FM, Loneragan JF (1972) Zinc absorption by wheat seedlings and the nature of its inhibition by alkaline earth cations. J Exp Bot 23:552–560CrossRefGoogle Scholar
  11. Clark C, Holland P, Smith G (1986) Chemical composition of bleeding xylem sap from kiwifruit vines. Ann Bot 58:353–362Google Scholar
  12. Cohen CK, Garvin DF, Kochian LV (2004) Kinetic properties of a micronutrient transporter from Pisum sativum indicate a primary function in Fe uptake from the soil. Planta 218:784–792CrossRefPubMedGoogle Scholar
  13. Coleman JE (1992) Zinc proteins: enzymes, storage proteins, transcription factors, and replication proteins. Annu Rev Biochem 61:897–946CrossRefPubMedGoogle Scholar
  14. Das P, Samantaray S, Rout GR (1997) Studies on cadmium toxicity in plants: a review. Environ Pollut 98:29–36CrossRefPubMedGoogle Scholar
  15. DiDonato RJ Jr, Roberts LA, Sanderson T, Eisley RB, Walker EL (2004) Arabidopsis Yellow Stripe-Like2 (YSL2): a metal-regulated gene encoding a plasma membrane transporter of nicotianamine-metal complexes. Plant J 39:403–414CrossRefPubMedGoogle Scholar
  16. Eide D, Kaplan S, Jordan I, Sipe D, Kaplan J (1992) Regulation of iron uptake in Saccharomyces cerevisiae. The ferrireductase and Fe(II) transporter are regulated independently. J Biol Chem 267:20774–20781PubMedGoogle Scholar
  17. Eide D, Broderius M, Fett J, Guerinot ML (1996) A novel iron-regulated metal transporter from plants identified by functional expression in yeast. PNAS 93:5624–5628CrossRefPubMedGoogle Scholar
  18. Eng BH, Guerinot ML, Eide D, Saier J (1998) Sequence analyses and phylogenetic characterization of the zip family of metal ion transport proteins. J Membr Biol 166:1–7CrossRefPubMedGoogle Scholar
  19. Freedman JH, Pickart L, Weinstein B, Mims WB, Peisach J (1982) Structure of the Glycyl-L-histidyl-L-lysine–copper(II) complex in solution. Biochemistry 21:4540–4544CrossRefPubMedGoogle Scholar
  20. Gietz DR, Schiestl RH (1991) Applications of high efficiency lithium acetate transformation of intact yeast cells using single-stranded nucleic acids as carrier. Yeast 7:253–263CrossRefPubMedGoogle Scholar
  21. Giordano PM, Noggle CJ, Mortvedt JJ (1974) Zinc uptake by rice, as affected by metabolic inhibitors and competing cations. Plant Soil 41:637–646CrossRefGoogle Scholar
  22. Grotz N, Fox T, Connolly E, Park W, Guerinot ML, Eide D (1998) Identification of a family of zinc transporter genes from Arabidopsis that respond to zinc deficiency. Proc Natl Acad Sci USA 95:7220–7224CrossRefPubMedGoogle Scholar
  23. Guerinot ML (2000) The ZIP family of metal transporters. Biochimica et Biophysica Acta-Biomembranes 1465:190–198CrossRefGoogle Scholar
  24. Hacisalihoglu G, Hart JJ, Kochian LV (2001) High- and low-affinity zinc transport systems and their possible role in zinc efficiency in bread wheat. Plant Physiol 125:456–463CrossRefPubMedGoogle Scholar
  25. Hall JL, Williams LE (2003) Transition metal transporters in plants. J Exp Bot 54:2601–2613CrossRefPubMedGoogle Scholar
  26. Haney CJ, Grass G, Franke S, Rensing C (2005) New developments in the understanding of the cation diffusion facilitator family. J Ind Microbiol Biotechnol 32:215–226CrossRefPubMedGoogle Scholar
  27. Hawf LR, Schmid WE (1967) Uptake and translocation of zinc by intact plants. Plant Soil 27:249–260CrossRefGoogle Scholar
  28. Hocking P (1980) The composition of phloem exudate and xylem sap from tree tobacco (Nicotiana glauca Grah.). Ann Bot 45:633–643Google Scholar
  29. Hocking P, Pate J, Atkins C, Sharkey P (1978) Diurnal patterns of transport and accumulation of minerals in fruiting plants of Lupinus angustifolius L. Ann Bot 42:1277–1290Google Scholar
  30. Kausar MA, Chaudhry FM, Rashid A, Latif A, Alam SM (1976) Micronutrient availability to cereals from calcareous soils. Plant Soil 45:397–410CrossRefGoogle Scholar
  31. Kochian LV (1991) Mechanisms of micronutrient uptake and translocation in plants. In: Mortvedt JJ, Cox FR, Shuman LM, Welch RM (eds) Micronutrients in agriculture. Soil Science Society of America Inc, Madison, pp 229–296Google Scholar
  32. Korenkov V, Hirschi K, Crutchfield JD, Wagner GJ (2007) Enhancing tonoplast Cd/H antiport activity increases Cd, Zn, and Mn tolerance, and impacts root/shoot Cd partitioning in Nicotiana tabacum L. Planta 226:1379–1387CrossRefPubMedGoogle Scholar
  33. Korshunova YO, Eide D, Gregg Clark W, Lou Guerinot M, Pakrasi HB (1999) The IRT1 protein from Arabidopsis thaliana is a metal transporter with a broad substrate range. Plant Mol Biol 40:37–44CrossRefPubMedGoogle Scholar
  34. Lipscomb WN, Strater N (1996) Recent advances in zinc enzymology. Chem Rev 96:2375–2434CrossRefPubMedGoogle Scholar
  35. López-Millán A-F, Ellis DR, Grusak MA (2004) Identification and characterization of several new members of the zip family of metal ion transporters in medicago truncatula. Plant Mol Biol 54:583–596CrossRefPubMedGoogle Scholar
  36. McCall KA, Huang C, Fierke CA (2000) Function and mechanism of zinc metalloenzymes. J Nutr 130:1437S–1446SPubMedGoogle Scholar
  37. Mills RF, Krijger GC, Baccarini PJ, Hall JL, Williams LE (2003) Functional expression of AtHMA4, a P-1B-type ATPase of the Zn/Co/Cd/Pb subclass. Plant J 35:164–176CrossRefPubMedGoogle Scholar
  38. Minet M, Dufour ME, Lacroute F (1992) Complementation of Saccharomyces cerevisiae auxotrophic mutants by Arabidopsis thaliana cDNAs. Plant J 2:417–422PubMedGoogle Scholar
  39. Moreau S, Thomson RM, Kaiser BN, Trevaskis B, Guerinot ML, Udvardi MK, Puppo A, Day DA (2002) GmZIP1 encodes a symbiosis-specific zinc transporter in soybean. J Biol Chem 277:4738–4746CrossRefPubMedGoogle Scholar
  40. Ooi CE, Rabinovich E, Dancis A, Bonifacino JS, Klausner RD (1996) Copper-dependent degradation of the Saccharomyces cerevisiae plasma membrane copper transporter Ctr1p in the apparent absence of endocytosis. EMBO J 15:3515–3523PubMedGoogle Scholar
  41. Orfei M, Alcaro MC, Marcon G, Chelli M, Ginanneschi M, Kozlowski H, Brasun J, Messori L (2003) Modeling of copper(II) sites in proteins based on histidyl and glycyl residues. J.Inorg.Biochem. 97:299–307CrossRefPubMedGoogle Scholar
  42. Pence NS, Larsen PB, Ebbs SD, Letham DLD, Lasat MM, Garvin DF, Eide D, Kochian LV (2000) The molecular physiology of heavy metal transport in the Zn/Cd hyperaccumulator Thlaspi caerulescens. Proc Natl Acad Sci USA 97:4956–4960CrossRefPubMedGoogle Scholar
  43. Petris MJ, Smith K, Lee J, Thiele DJ (2003) Copper-stimulated endocytosis and degradation of the human copper transporter, hCtr1. J Biol Chem 278:9639–9646CrossRefPubMedGoogle Scholar
  44. Sukkariyah BF, Evanylo G, Zelazny L, Chaney RL (2005) Cadmium, copper, nickel, and zinc availability in a biosolids-amended piedmont soil years after application. J Environ Qual 34:2255–2262CrossRefPubMedGoogle Scholar
  45. Vallee BL, Auld DS (1990) Zinc coordination, function, and structure of zinc enzymes and other proteins. Biochemistry 29:5647–5659CrossRefPubMedGoogle Scholar
  46. Wagner GJ (1993) Accumulation of cadmium in crop plants and its consequences to human health. In: Donald LS (ed) Advances in Agronomy. Academic Press, San Diego, pp 173–212Google Scholar
  47. Weast R (1976) CRC Handbook of Chemistry and Physics. CRC Press, ClevelandGoogle Scholar
  48. White MC, Decker AM, Chaney RL (1981) Metal Complexation in Xylem Fluid: I. Chemical Composition of Tomato And Soybean Stem Exudate. Plant Physiol 67:292–300CrossRefPubMedGoogle Scholar
  49. Zhao H, Eide D (1996a) The yeast ZRT1 gene encodes the zinc transporter protein of a high-affinity uptake system induced by zinc limitation. PNAS 93:2454–2458CrossRefPubMedGoogle Scholar
  50. Zhao H, Eide D (1996b) The ZRT2 gene encodes the low affinity zinc transporter in Saccharomyces cerevisiae. J Biol Chem 271:23203–23210CrossRefPubMedGoogle Scholar

Copyright information

© U.S. Government 2010

Authors and Affiliations

  • Brian W. Stephens
    • 1
    • 2
  • Douglas R. Cook
    • 2
  • Michael A. Grusak
    • 1
    Email author
  1. 1.Department of Pediatrics, USDA-ARS Children’s Nutrition Research CenterBaylor College of MedicineHoustonUSA
  2. 2.Department of Plant Pathology, Plant Biology Graduate GroupUniversity of California-DavisDavisUSA

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