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EMBO J. Jun 15, 1999; 18(12): 3325–3333.
PMCID: PMC1171413

Tolerance to toxic metals by a gene family of phytochelatin synthases from plants and yeast.

Abstract

Phytochelatins play major roles in metal detoxification in plants and fungi. However, genes encoding phytochelatin synthases have not yet been identified. By screening for plant genes mediating metal tolerance we identified a wheat cDNA, TaPCS1, whose expression in Saccharomyces cerevisiae results in a dramatic increase in cadmium tolerance. TaPCS1 encodes a protein of approximately 55 kDa with no similarity to proteins of known function. We identified homologs of this new gene family from Arabidopsis thaliana, Schizosaccharomyces pombe, and interestingly also Caenorhabditis elegans. The Arabidopsis and S.pombe genes were also demonstrated to confer substantial increases in metal tolerance in yeast. PCS-expressing cells accumulate more Cd2+ than controls. PCS expression mediates Cd2+ tolerance even in yeast mutants that are either deficient in vacuolar acidification or impaired in vacuolar biogenesis. PCS-induced metal resistance is lost upon exposure to an inhibitor of glutathione biosynthesis, a process necessary for phytochelatin formation. Schizosaccharomyces pombe cells disrupted in the PCS gene exhibit hypersensitivity to Cd2+ and Cu2+ and are unable to synthesize phytochelatins upon Cd2+ exposure as determined by HPLC analysis. Saccharomyces cerevisiae cells expressing PCS produce phytochelatins. Moreover, the recombinant purified S.pombe PCS protein displays phytochelatin synthase activity. These data demonstrate that PCS genes encode phytochelatin synthases and mediate metal detoxification in eukaryotes.

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Selected References

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  • Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. [PubMed]
  • 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 U S A. 1992 May 1;89(9):3736–3740. [PMC free article] [PubMed]
  • Clemens S, Antosiewicz DM, Ward JM, Schachtman DP, Schroeder JI. The plant cDNA LCT1 mediates the uptake of calcium and cadmium in yeast. Proc Natl Acad Sci U S A. 1998 Sep 29;95(20):12043–12048. [PMC free article] [PubMed]
  • Fahey RC, Newton GL. Determination of low-molecular-weight thiols using monobromobimane fluorescent labeling and high-performance liquid chromatography. Methods Enzymol. 1987;143:85–96. [PubMed]
  • Goyer RA. Toxic and essential metal interactions. Annu Rev Nutr. 1997;17:37–50. [PubMed]
  • Grill E, Winnacker EL, Zenk MH. Phytochelatins: the principal heavy-metal complexing peptides of higher plants. Science. 1985 Nov 8;230(4726):674–676. [PubMed]
  • Grill E, Löffler S, Winnacker EL, Zenk MH. Phytochelatins, the heavy-metal-binding peptides of plants, are synthesized from glutathione by a specific gamma-glutamylcysteine dipeptidyl transpeptidase (phytochelatin synthase). Proc Natl Acad Sci U S A. 1989 Sep;86(18):6838–6842. [PMC free article] [PubMed]
  • Hayashi Y, Nakagawa CW, Mutoh N, Isobe M, Goto T. Two pathways in the biosynthesis of cadystins (gamma EC)nG in the cell-free system of the fission yeast. Biochem Cell Biol. 1991 Feb-Mar;69(2-3):115–121. [PubMed]
  • Ho MN, Hill KJ, Lindorfer MA, Stevens TH. Isolation of vacuolar membrane H(+)-ATPase-deficient yeast mutants; the VMA5 and VMA4 genes are essential for assembly and activity of the vacuolar H(+)-ATPase. J Biol Chem. 1993 Jan 5;268(1):221–227. [PubMed]
  • Howden R, Goldsbrough PB, Andersen CR, Cobbett CS. Cadmium-sensitive, cad1 mutants of Arabidopsis thaliana are phytochelatin deficient. Plant Physiol. 1995 Apr;107(4):1059–1066. [PMC free article] [PubMed]
  • Kneer R, Kutchan TM, Hochberger A, Zenk MH. Saccharomyces cerevisiae and Neurospora crassa contain heavy metal sequestering phytochelatin. Arch Microbiol. 1992;157(4):305–310. [PubMed]
  • Kramer U, Smith RD, Wenzel WW, Raskin I, Salt DE. The Role of Metal Transport and Tolerance in Nickel Hyperaccumulation by Thlaspi goesingense Halacsy. Plant Physiol. 1997 Dec;115(4):1641–1650. [PMC free article] [PubMed]
  • Lemen RA, Lee JS, Wagoner JK, Blejer HP. Cancer mortality among cadmium production workers. Ann N Y Acad Sci. 1976;271:273–279. [PubMed]
  • Mewes HW, Albermann K, Bähr M, Frishman D, Gleissner A, Hani J, Heumann K, Kleine K, Maierl A, Oliver SG, et al. Overview of the yeast genome. Nature. 1997 May 29;387(6632 Suppl):7–65. [PubMed]
  • Moreno S, Klar A, Nurse P. Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol. 1991;194:795–823. [PubMed]
  • Mutoh N, Hayashi Y. Isolation of mutants of Schizosaccharomyces pombe unable to synthesize cadystin, small cadmium-binding peptides. Biochem Biophys Res Commun. 1988 Feb 29;151(1):32–39. [PubMed]
  • Okazaki K, Okazaki N, Kume K, Jinno S, Tanaka K, Okayama H. High-frequency transformation method and library transducing vectors for cloning mammalian cDNAs by trans-complementation of Schizosaccharomyces pombe. Nucleic Acids Res. 1990 Nov 25;18(22):6485–6489. [PMC free article] [PubMed]
  • Ortiz DF, Kreppel L, Speiser DM, Scheel G, McDonald G, Ow DW. Heavy metal tolerance in the fission yeast requires an ATP-binding cassette-type vacuolar membrane transporter. EMBO J. 1992 Oct;11(10):3491–3499. [PMC free article] [PubMed]
  • Ortiz DF, Ruscitti T, McCue KF, Ow DW. Transport of metal-binding peptides by HMT1, a fission yeast ABC-type vacuolar membrane protein. J Biol Chem. 1995 Mar 3;270(9):4721–4728. [PubMed]
  • Raskin I, I, Smith RD, Salt DE. Phytoremediation of metals: using plants to remove pollutants from the environment. Curr Opin Biotechnol. 1997 Apr 1;8(2):221–226. [PubMed]
  • Rauser WE. Phytochelatins. Annu Rev Biochem. 1990;59:61–86. [PubMed]
  • Rauser WE. Phytochelatins and related peptides. Structure, biosynthesis, and function. Plant Physiol. 1995 Dec;109(4):1141–1149. [PMC free article] [PubMed]
  • Rea Philip A, Li Ze-Sheng, Lu Yu-Ping, Drozdowicz Yolanda M, Martinoia Enrico. FROM VACUOLAR GS-X PUMPS TO MULTISPECIFIC ABC TRANSPORTERS. Annu Rev Plant Physiol Plant Mol Biol. 1998 Jun;49(NaN):727–760. [PubMed]
  • Robinson JS, Graham TR, Emr SD. A putative zinc finger protein, Saccharomyces cerevisiae Vps18p, affects late Golgi functions required for vacuolar protein sorting and efficient alpha-factor prohormone maturation. Mol Cell Biol. 1991 Dec;11(12):5813–5824. [PMC free article] [PubMed]
  • Rodríguez-Navarro A, Ramos J. Dual system for potassium transport in Saccharomyces cerevisiae. J Bacteriol. 1984 Sep;159(3):940–945. [PMC free article] [PubMed]
  • Salt DE, Wagner GJ. Cadmium transport across tonoplast of vesicles from oat roots. Evidence for a Cd2+/H+ antiport activity. J Biol Chem. 1993 Jun 15;268(17):12297–12302. [PubMed]
  • Schachtman DP, Schroeder JI. Structure and transport mechanism of a high-affinity potassium uptake transporter from higher plants. Nature. 1994 Aug 25;370(6491):655–658. [PubMed]
  • Silver S, Phung LT. Bacterial heavy metal resistance: new surprises. Annu Rev Microbiol. 1996;50:753–789. [PubMed]
  • Stohs SJ, Bagchi D. Oxidative mechanisms in the toxicity of metal ions. Free Radic Biol Med. 1995 Feb;18(2):321–336. [PubMed]
  • Thompson JD, Higgins DG, Gibson TJ. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994 Nov 11;22(22):4673–4680. [PMC free article] [PubMed]
  • Thumann J, Grill E, Winnacker EL, Zenk MH. Reactivation of metal-requiring apoenzymes by phytochelatin-metal complexes. FEBS Lett. 1991 Jun 17;284(1):66–69. [PubMed]
  • Zenk MH. Heavy metal detoxification in higher plants--a review. Gene. 1996 Nov 7;179(1):21–30. [PubMed]

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