The IRT1 protein from Arabidopsis thaliana is a metal transporter with a broad substrate range Abstract
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 References
Andersson B, Styring S: Photosystem II: molecular organization, function and acclimation. Curr Top Bioenerg 16: 1–81 (1991).
Anderson JA, Huprikar SS, Kochian LV, Lucas WJ, Gaber RF: Functional expression of a probable
potassium channel in
. Proc Natl Acad Sci USA 89: 3736–3740 (1992).
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).
Bartsevich VV, Pakrasi HB: Manganese transport in the cyanobacterium
sp. PCC 6803. J Biol Chem 271: 26057–26061 (1996).
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).
Belouchi AM, Kwan T, Gros P: Cloning and characterization of the
, a new family of membrane proteins possibly implicated in the transport of metal ions. Plant Mol Biol 33: 1085–1092 (1997).
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).
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).
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).
Dancis A, Yuan DS, Haile D, Askwith C, Eide D, Moehle C, Kaplan J, Klausner DR: Molecular characterization of a copper transport protein in
: an unexpected role for copper in iron transport. Cell 76: 393–402 (1994).
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).
Feinberg AP, Vogelstein B: A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 137: 266–267 (1984).
Frausto da Silva JJR, Williams RJP: The Biological Chemistry of the Elements: The Inorganic Chemistry of Life. Oxford University Press, New York (1991).
Glerum DM, Shtanko A, Tzagoloff A: Characterization of
, a yeast gene involved in copper metabolism and assembly of cytochrome oxidase. J Biol Chem 271: 14504–14509 (1996).
Grotz N, Fox T, Connolly E, Park W, Guerinot ML, Eide D: Identification of a family of zinc transporter genes from
that respond to zinc deficiency. Proc Natl Acad Sci USA 95: 7220–7224 (1998).
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).
Kampfenkel K, Kushnir S, Babiychuk E, Inze D, Van Montagu MV:Molecular characterization of a putative
copper transporter and its yeast homologue. J Biol Chem 270: 28479–28486 (1995).
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).
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).
Lin S-J, Culotta Cizeweski V: The
encodes a small metal homeostasis factor that protects cells against reactive oxygen toxicity. Proc Natl Acad Sci USA 92: 3784–3788 (1995).
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).
Marschner H: Mineral Nutrition of Higher Plants. Academic Press, London (1986).
Minet M, Dufour M-E, Lacroute F: Complementation of
auxotrophic mutants by
cDNAs. Plant J 2: 417–422 (1992).
Murashige T, Skoog F: A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497 (1962).
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
. Plant Physiol 103: 553–564 (1993).
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).
Rodecap KD, Tingey DT, Lee EH: Iron nutrition influence on cadmium accumulation by
(L.) Heynh. J Environ Qual 23: 239–246 (1994).
Sambrook J, Fritsch E, Maniatis T: Molecular Cloning: A LaboratoryManual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).
Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).
Schachtman DP, Schroeder JI: Structure and transport mechanism of a high-affinity potassium uptake transporter from higher plants. Nature 370: 655–658 (1994).
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).
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).
Sherman F, Fink GR, Hicks JB: Laboratory course manual for methods in yeast genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1986).
Strathern JN, Higgins DR: Recovery of plasmids from yeast into
shuttle vectors. Meth Enzymol 194: 319–328 (1991).
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).
Verwoerd TC, Dekker BMM, Hoekema A: A small-scale procedure for the rapid isolation of plant RNAs. Nucl Acids Res 17: 2362 (1989).
Welch RM, Norvell WA, Schaefer SC, Shaff JE, Kochian LV: Induction of iron(III) and copper(II) reduction in pea (
) 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).
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).
Zhao H, Eide D: The
gene encodes the low affinity zinc transporter in
J Biol Chem 271: 23203–23210 (1996).
Zhao H, Eide D: The yeast
gene encodes the zinc transporter protein of high-affinity uptake system induced by zinc limitation. Proc Natl Acad Sci USA 93: 2454–2458 (1996).
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