Plant Molecular Biology

, Volume 58, Issue 6, pp 839–855 | Cite as

The Plant MT1 Metallothioneins are Stabilized by Binding Cadmiums and are Required for Cadmium Tolerance and Accumulation

  • Anne Marie Zimeri
  • Om Parkash Dhankher
  • Bonnie McCaig
  • Richard B. Meagher
Article

Abstract

The small Arabidopsis genome contains nine metallothionein-like (MT) sequences with classic, cysteine-rich domains separated by spacer sequences, quite unlike the small conserved MT families found vertebrate genomes. Phylogenetic analysis revealed four ancient and divergent classes of plant MTs that predate the monocot–dicot divergence. A distinct cysteine spacing pattern suggested differential metal ion specificity for each class. The in vivo stability of representatives of the four classes of plant MT proteins and a mouse MT2 control expressed in E. coli were enhanced by cadmium (Cd). Particular MTs were also stabilized by arsenic (As), copper (Cu), and or zinc (Zn). To understand why plants have such a diversity of MT sequences, the Arabidopsis MT1 class, comprised of three genes, MT1a, MT1b, and MT1c, was characterized in more detail in plants. MT1 family transcripts were knocked down to less than 5–10% of wild-type levels in Arabidopsis by expression of a RNA interference (RNAi) construct. The MT1 knockdown plant lines were all hypersensitive to Cd and accumulated several fold lower levels of As, Cd, and Zn than wildtype, while Cu and Fe levels were unaffected. The ancient class of MT1 protein sequences may be preserved in plant genomes, because it has distinct metal-binding properties, confers tolerance to cadmium, and can assist with zinc homeostasis.

Keywords

cysteine spacing pattern evolution metal accumulation RNA interference thiol reactivity 

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References

  1. Adriano, D.C. 1986Trace elements in the Terrestrial EnvironmentSpringer VerlagNew YorkCrossRefGoogle Scholar
  2. An, G., Ebert, P.R., Mitra, A., Ha, S.B. 1988Binary vectorsGelvin, S.B.Schilperoot, R.A. eds. Plant Molecular BiologyMartinus NijhoffAmsterdam119Google Scholar
  3. An, Y.-Q., McDowell, J.M., Huang, S., McKinney, E.C., Chambliss, S., Meagher, R.B. 1996Strong, constitutive expression of the Arabidopsis ACT2/ACT8 actin subclass in vegetative tissuesPlant J.10107121CrossRefGoogle Scholar
  4. Arabidopsis Genome Initiative2000Analysis of the genome sequence of the flowering plant Arabidopsis thalianaNature408796815CrossRefGoogle Scholar
  5. Arnold,  1991Engineered metal-binding proteins: purification to protein foldingScience25217961797CrossRefGoogle Scholar
  6. Arseniev, A., Schultze, P., Worgotter, E., Braun, W., Wagner, G., Vasak, M., Kagi, J.H., Wuthrich, K. 1988Three-dimensional structure of rabbit liver (Cd7) metallothionein−2a in aqueous solution determined by nuclear magnetic resonanceJ. Mol. Biol.201637657CrossRefGoogle Scholar
  7. Baryla, A., Carrier, P., Franck, F., Coulomb, C., Sahut, C., Havaux, M. 2001Leaf chlorosis in oilseed rape plants (Brassica napus) grown on cadmium-polluted soil: causes and consequences for photosynthesis and growthPlanta212696709CrossRefGoogle Scholar
  8. Bredenberg, J., Nilsson,  2002Conformational states of the glucocorticoid receptor DNA-binding domain from molecular dynamics simulationsProteins492436CrossRefGoogle Scholar
  9. Buhtz, A., Kolasa, A., Arlt, K., Walz, C., Kehr, J. 2004Xylem sap protein composition is conserved among different plant speciesPlanta219610618CrossRefGoogle Scholar
  10. Cha, J., Pedersen, M.V., Auld, D.S. 1996Metal and pH dependence of heptapeptide catalysis by human matrilysinBiochemistry351583115838CrossRefGoogle Scholar
  11. Cobbett, C., Goldsbrough, P. 2002Phytochelatins and metallothioneins: roles in heavy metal detoxification and homeostasisAnnu. Rev. Plant. Biol.53159182CrossRefGoogle Scholar
  12. Dhankher, O.P., Li, Y., Rosen, B.P., Shi, J., Salt, D., Senecoff, J.F., Sashti, N.A., Meagher, R.B. 2002Engineering tolerance and hyperaccumulation of arsenic in plants by combining arsenate reductase and gamma-glutamylcysteine synthetase expressionNat. Biotechnol.2011401145CrossRefGoogle Scholar
  13. Dhankher, O.P., Rosen, B.P., Fuhrman, M., Meagher, R.B. 2003Increased cadmium tolerance and accumulation by plants expressing bacterial arsenate reductaseNew Phytol.159431441CrossRefGoogle Scholar
  14. Ejnik, J., Robinson, J., Zhu, J., Forsterling, H., Shaw, C.F., Petering, D.H. 2002Folding pathway of apo-metallothionein induced by Zn2+, Cd2+ and Co2+J. Inorg. Biochem.88144152CrossRefGoogle Scholar
  15. Evans, K.M., Gatehouse, J.A., Lindsay, W.P., Shi, J., Tommey, A.M., Robinson, N.J. 1992Expression of the pea metallothionein-like gene PsMTA in Escherichia coli and Arabidopsis thaliana and analysis of trace metal ion accumulation: implications for PsMTA functionPlant Mol. Biol.2010191028CrossRefGoogle Scholar
  16. Fern, R., Black, J.A., Ransom, B.R., Waxman, S.G. 1996Cd(2+)-induced injury in CNS white matterJ. Neurophysiol.7632643273CrossRefGoogle Scholar
  17. Gilliland, L.U., Pawloski, L., Kandasamy, M.K., Meagher, R.B. 2002The Arabidopsis actin isovariant, ACT7, plays a vital role in germintion and root growthPlant J.33319328CrossRefGoogle Scholar
  18. Guo, W., Bundithya, W., Goldsbrough, P. 2003Characterization of the Arabidopsis metallothionein gene family: tissue-specific expression and induction during senescence and in response to copperNew Phytol.159369381CrossRefGoogle Scholar
  19. Hamer, D.H. 1986MetallothioneinAnn. Rev. Biochem.55913951CrossRefGoogle Scholar
  20. Hannon, G.J. 2002RNA interferenceNature418244251CrossRefGoogle Scholar
  21. Hasler, D.W., Jensen, L.T., Zerbe, O., Winge, D.R., Vasak, M. 2000Effect of the two conserved prolines of human growth inhibitory factor (metallothionein−3) on its biological activity and structure fluctuation: comparison with a mutant proteinBiochemistry391456714575CrossRefGoogle Scholar
  22. Howden, R., Goldsbrough, P.B., Andersen, C.R., Cobbett, C.S. 1995Cadmium-sensitive cad1 mutants of Arabidopsis thaliana are phytochelatin deficientPlant Physiol.10710591066CrossRefGoogle Scholar
  23. Jacob, C., Maret, W., Vallee, B.L. 1998aControl of zinc transfer between thionein, metallothionein, and zinc proteinsProc. Natl. Acad. Sci. USA9534893494CrossRefGoogle Scholar
  24. Jacob, C., Maret, W., Vallee, B.L. 1998bEbselen, a selenium-containing redox drug, releases zinc from metallothioneinBiochem. Biophys. Res. Commun.248569573CrossRefGoogle Scholar
  25. Jiang, L.J., Maret, W., Vallee, B.L. 1998The glutathione redox couple modulates zinc transfer from metallothionein to zinc-depleted sorbitol dehydrogenaseProc. Natl. Acad. Sci. USA9534833488CrossRefGoogle Scholar
  26. Jiang, L.J., Vasak, M., Vallee, B.L., Maret, W. 2000Zinc transfer potentials of the alpha - and beta-clusters of metallothionein are affected by domain interactions in the whole moleculeProc. Natl. Acad. Sci. USA9725032508CrossRefGoogle Scholar
  27. Kagi, J.H., Schaffer, A. 1988Biochemistry of metallothioneinBiochemistry2785098515CrossRefGoogle Scholar
  28. Kandasamy, M.K., McKinney, E.C., Meagher, R.B. 2002Plant profilin isovariants are distinctly regulated in vegetative and reproductive tissuesCell Motil. Cytoskeleton522232CrossRefGoogle Scholar
  29. Kenrick, P., Crane, P.R. 1997The origin and early evolution of plants on landNature3893339CrossRefGoogle Scholar
  30. Kille, P., Winge, D.R., Harwood, J.L., Kay, J. 1991A plant metallothionein produced in E. coliFEBS Lett.295171175CrossRefGoogle Scholar
  31. Klaassen, C.D., Liu, J., Choudhuri, S. 1999Metallothionein: an intracellular protein to protect against cadmium toxicityAnnu. Rev. Pharmacol. Toxicol.39267294CrossRefGoogle Scholar
  32. Lefebvre, D.D., Miki, B.L., Laliberte, J.F. 1987Mammalian metallothionein functions in plantsBiotechnology510531056Google Scholar
  33. Li, W.-H., Wu, C.-I., Luo, C.-C. 1986A new method for estimating synonymous and nonsynonymous rates of nucleotide substitution considering the relative likelihood of nucleotide and codon usageMol. Biol. Evol.2150174Google Scholar
  34. Li, Y., Cockburn, W., Kilpatrick, J., Whitelam, G.C. 2000Cytoplasmic expression of a soluble synthetic mammalian metallothionein-alpha domain in Escherichia coli. Enhanced tolerance and accumulation of cadmiumMol. Biotechnol.16211219CrossRefGoogle Scholar
  35. Liu, J., Liu, Y., Habeebu, S.S., Klaassen, C.D. 1999aMetallothionein-null mice are highly susceptible to the hematotoxic and immunotoxic effects of chronic CdCl2 exposureToxicol. Appl. Pharmacol.15998108CrossRefGoogle Scholar
  36. Liu, Y., Liu, J., Habeebu, S.S., Klaassen, C.D. 1999bMetallothionein protects against the nephrotoxicity produced by chronic CdMT exposureToxicol. Sci.50221227CrossRefGoogle Scholar
  37. Luisi, B.F., Xu, W.X., Otwinowski, Z., Freedman, L.P., Yamamoto, K.R., Sigler, P.B. 1991Crystallographic analysis of the interaction of the glucocorticoid receptor with DNANature352497505CrossRefGoogle Scholar
  38. Magonet, E., Hayen, P., Delforge, D., Delaive, E., Remacle, J. 1992Importance of the structural zinc atom for the stability of yeast alcohol dehydrogenaseBiochem. J.287361365CrossRefGoogle Scholar
  39. Maret, W. 2000The function of zinc metallothionein: a link between cellular zinc and redox stateJ. Nutr.1301455S1458SCrossRefGoogle Scholar
  40. Mayo, K.E., Palmiter, R.D. 1982Glucocorticoid regulation of the mouse metallothionein I gene is selectively lost following amplification of the geneJ. Biol. Chem.25730613067PubMedGoogle Scholar
  41. Mayo, K.E., Warren, R., Palmiter, R.D. 1982The mouse metallothionein-I gene is transcriptionally regulated by cadmium following transfection into human or mouse cellsCell2999108CrossRefGoogle Scholar
  42. Miccadei, S., Floridi, A. 1993Sites of inhibition of mitochondrial electron transport by cadmiumChem. Biol. Interact.8915967CrossRefGoogle Scholar
  43. Michalska, A.E., Choo, K.H. 1993Targeting and germ-line transmission of a null mutation at the metallothionein I and II loci in mouseProc. Natl. Acad. Sci. USA9080888092CrossRefGoogle Scholar
  44. Misra, R.R., Hochadel, J.F., Smith, G.T., Cook, J.C., Waalkes, M.P., Wink, D.A. 1996Evidence that nitric oxide enhances cadmium toxicity by displacing the metal from metallothioneinChem. Res. Toxicol.9326332CrossRefGoogle Scholar
  45. Misra, S., Gedamu, L. 1989Heavy metal tolerant transgenic Brassica napus L. and Nicotiana tabacum L. plantsTheor. Appl. Genet.78161168CrossRefGoogle Scholar
  46. Morris, C.A., Nicolaus, B., Sampson, V., Harwood, J.L., Kille, P. 1999Identification and characterization of a recombinant metallothionein protein from a marine alga, Fucus vesiculosusBiochem. J.338553560CrossRefGoogle Scholar
  47. Murashige, T., Skoog, F. 1962A revised medium for rapid growth and bioassays with tobacco tissue culturePlant Physiol.15473497CrossRefGoogle Scholar
  48. Murphy, A., Zhou, J., Goldsbrough, P.B., Taiz, L. 1997Purification and immunological identification of metallothioneins 1 and 2 from Arabidopsis thalianaPlant Physiol.11312931301CrossRefGoogle Scholar
  49. O’Hara, E.B., Chekanova, J.A., Ingle, C.A., Kushner, Z.R., Peters, E., Kushner, S.R. 1995Polyadenylylation helps regulate mRNA decay in Escherichia coliProc. Natl. Acad. Sci. USA9218071811CrossRefGoogle Scholar
  50. Palmiter, R.D. 1998The elusive function of metallothioneinsProc. Natl. Acad. Sci. USA9584288430CrossRefGoogle Scholar
  51. Rae, T.D., Schmidt, P.J., Pufahl, R.A., Culotta, V.C., O’Halloran, T.V. 1999Undetectable intracellular free copper: the requirement of a copper chaperone for superoxide dismutaseScience284805808CrossRefGoogle Scholar
  52. Robbins, A.H., McRee, D.E., Williamson, M., Collett, S.A., Xuong, N.H., Furey, W.F., Wang, B.C., Stout, C.D. 1991Refined crystal structure of Cd, Zn metallothionein at 2.0 Å resolutionJ. Mol. Biol.22112691293PubMedGoogle Scholar
  53. Robinson, N.J., Tommey, A.M., Kuske, C., Jackson, P.J. 1993Plant metallothioneinsBiochem. J.295110CrossRefGoogle Scholar
  54. Roschitzki, B., Vasak, M. 2003Redox labile site in a Zn4 cluster of Cu4, Zn4-metallothionein−3Biochemistry4298229828CrossRefGoogle Scholar
  55. Saitou, N., Nei, M 1987The neighbour-joining method: a new method for reconstructing phylogenetic treesMol. Biol. Evol.4406425PubMedGoogle Scholar
  56. Sanderson, M.J., Doyle, J.A. 2001Sources of error and confidence intervals in estimating the age of angiosperms from RBCL and 18S rDNA dataAm. J. Bot.8814991516CrossRefGoogle Scholar
  57. Schneider, G., Eklund, H., Cedergren-Zeppezauer, E., Zeppezauer, M. 1983Structure of the complex of active site metal-depleted horse liver alcohol dehydrogenase and NADHEMBO J.2685689PubMedPubMedCentralGoogle Scholar
  58. Shirley, B.W., Meagher, R.B. 1990A potential role for RNA turnover in the light regulation of plant gene expression: ribulose−1,5-bisphosphate carboxylase small subunit in soybeanNuc. Acids Res.1833773385CrossRefGoogle Scholar
  59. Swofford, D. 1998PAUP: Phylogenetic analysis using parsimony. Version 4Sinauer AssociatesSunderland, MassachusettsGoogle Scholar
  60. Ubbink, M., Lian, L.Y., Modi, S., Evans, P.A. and Bendall, D.S. 1996. Analysis of the 1H-NMR chemical shifts of Cu(I)-, Cu(II)- and Cd-substituted pea plastocyanin Metaldependent. 242: 132-147.Google Scholar
  61. Wei, D.Y., Andrews, G.K. 1988Molecular cloning of chicken metallothionein. Deduction of the complete amino acid sequence and analysis of expression using cloned cDNANucleic Acids Res.16537553CrossRefGoogle Scholar
  62. Yang, Y., Maret, W., Vallee, B.L. 2001Differential fluorescence labeling of cysteinyl clusters uncovers high tissue levels of thioneinProc. Natl. Acad. Sci. USA9855565559CrossRefGoogle Scholar
  63. Ye, B., Maret, W., Vallee, B.L. 2001Zinc metallothionein imported into liver mitochondria modulates respirationProc. Natl. Acad. Sci. USA9823172322CrossRefGoogle Scholar
  64. Zambryski, P., Crawford, K. 2000Plasmodesmata: gatekeepers for cell-to-cell transport of developmental signals in plantsAnnu. Rev. Cell Dev. Biol.16393421CrossRefGoogle Scholar
  65. Zangger, K., Oz, G., Otvos, J.D., Armitage, I.M. 1999Three-dimensional solution structure of mouse (Cd7)-metallothionein−1 by homonuclear and heteronuclear NMR spectroscopyProtein Sci.826302638CrossRefGoogle Scholar
  66. Zhou, J., Goldsbrough, P.B. 1995Structure, organization and expression of the metallothionein gene family in ArabidopsisMol. Gen. Genet.248318328CrossRefGoogle Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • Anne Marie Zimeri
    • 1
    • 2
  • Om Parkash Dhankher
    • 1
    • 3
  • Bonnie McCaig
    • 4
  • Richard B. Meagher
    • 1
  1. 1.Department of GeneticsUniversity of GeorgiaAthensUSA
  2. 2.Toxicology & Mycology Research UnitUSDA-ARS, Russell Research CenterAthensUSA
  3. 3.Department of Plant, Soil and Insect SciencesUniversity of MassachusettsAmherstUSA
  4. 4.D.O.E. Plant Research LaboratoriesMichigan State UniversityEast LansingUSA

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