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Cloning and expression of amino acid transporters from broad bean

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Abstract

This work describes the isolation of a full-length (VfAAP2) and three partial amino acid transporter genes (VfAAPa, VfAAPb, VfAAPc) from broad bean (Vicia faba L.). The function of VfAAP2 was tested by heterologous expression in a yeast mutant deficient in proline uptake. VfAAP2 mediates proton-dependent proline uptake with an apparent Km of about 1 mM. Analysis of substrate specificity by competition experiments showed that aromatic amino acids, neutral aliphatic acids and L-citrulline are the best competitors, whereas basic amino acids do not compete with proline. Northern analysis indicates that all VfAAPs exhibit different patterns of expression. VfAAP2 is most strongly expressed in the stem and at a lower level in sink leaves and pods. VfAAPa, VfAAPb and VfAAPc are most strongly expressed in the flowers, but their expression in the other organs varies.

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References

  • Altschul, S.F., Gish, W., Miller, W., Myers, E.W. and Lipman, D.J. 1990. Basic local alignment search tool. J Mol Biol 215: 403–410.

    Google Scholar 

  • Bennett, M.J., Marchant, A., Green, H.G., May, S.T., Ward, S.P., Millner, P.A., Walker, A.R., Schulz, B. and Feldmann, K.A. 1996. Arabidopsis AUX1 gene: a permease-like regulator of root gravitropism. Science 273: 948–950.

    Google Scholar 

  • Bick, J.A., Neelam, A., Hall, J.L. and Williams, L.E. 1998. Amino acid carriers of Ricinus communis expressed during seedling development: molecular cloning and expression analysis of two putative amino acid transporters, RcAAP1 and RcAAP2. Plant Mol. Biol. 36: 377–385.

    Google Scholar 

  • Boorer, K.J., Frommer, W.B., Bush, D.R., Kreman, M., Loo, D.D. and Wright, E.M. 1996. Kinetics and specificity of a HC/amino acid transporter from Arabidopsis thaliana. J. Biol. Chem. 271: 2213–2220.

    Google Scholar 

  • Boorer, K.J. and Fischer, W.N. 1997. Specificity and stoichiometry of the Arabidopsis HC/amino acid transporter AAP5. J. Biol. Chem. 272: 13040–13046.

    Google Scholar 

  • Bush, D. 1993. Proton-coupled sugar and amino acid transporters in plants. Annu. Rev. Plant Physiol. Plant Mol. Biol. 44: 513–542.

    Google Scholar 

  • Chang, H.C. and Bush, D.R. 1997. Topology of NAT2, a prototypical example of a new family of amino acid transporters. J Biol. Chem. 272: 30552–30557.

    Google Scholar 

  • Chen, L. and Bush, D.R. 1997. LHT1, a lysine and histidine specific amino acid transporter in Arabidopsis. Plant Physiol. 115: 1127–1134.

    Google Scholar 

  • Despeghel, J.P. 1981. Etude des mécanismes de l'absorption des acides aminés neutres par les tissus foliaires et de leur accumulation dans les nervures. Ph.D. thesis, University of Poitiers, France.

  • Despeghel, J.P. and Delrot, S. 1983. Energetics of amino acid uptake by Vicia faba leaf tissue. Plant Physiol 71: 1–6.

    Google Scholar 

  • Dohmen, R.J., Strasser, A.W.M., Höner, C.B. and Hollenberg, C.P. 1991. An efficient transformation procedure enabling long-term storage of competent cells of various yeast genera. Yeast 7: 691–692.

    Google Scholar 

  • Etherton, B. and Rubinstein, B. 1978. Evidence for amino acid-HC co-transport in oat coleoptiles. Plant Physiol. 61: 933–937.

    Google Scholar 

  • Fischer, W.N., Kwart, M., Hummel, S. and Frommer, W.B. 1995. Substrate specificity and expression profile of amino acid transporters (AAPs) in Arabidopsis. J. Biol. Chem. 270: 16315–16320.

    Google Scholar 

  • Fischer, W.N., André, B., Rentsch, D., Krolkiewicz, S., Tegeder, M., Breitkreuz, K. and Frommer, W.B. 1998. Amino acid transport in plants. Trends Plant Sci. 3: 188–195.

    Google Scholar 

  • Frommer, W.B., Hummel, S. and Riesmeier, J.W. 1993. Expression cloning in yeast of a cDNA encoding a broad specificity amino acid permease from Arabidopsis thaliana. Proc. Natl. Acad. Sci. USA 90: 5944–5948.

    Google Scholar 

  • Frommer, W.B., Hummel, S. and Rentsch, D. 1994. Cloning of an Arabidopsis histidine transporting protein related to nitrate and peptide transporters. FEBS Lett. 347: 185–189.

    Google Scholar 

  • Frommer, W.B., Hummel, S., Unseld, M. and Ninnemann, O. 1995. Seed and vascular expression of a high-affinity transporter for cationic amino acids in Arabidopsis. Proc. Natl. Acad. Sci. USA 92: 12036–12040.

    Google Scholar 

  • Girousse, C., Bournoville, R. and Bonnemain, J.L. 1996. Waterdeficit induced changes in concentrations in proline and some other amino acids in the phloem sap of alfalfa. Plant Physiol. 111: 109–113.

    Google Scholar 

  • Hirner, B., Fischer, W.N., Rentsch, D., Kwart, M. and Frommer, W.B. 1998. Developmental control of HC/amino acid permease gene expression during seed development of Arabidopsis. Plant J. 14: 535–544.

    Google Scholar 

  • Hoagland, D.R. and Snyder, W.C. 1933. Nutrition of strawberry plants under controlled conditions. Proc. Am. Soc. Hort. 30: 288–296.

    Google Scholar 

  • Hofmann, K. and Stoffel, W. 1993. Tmbase: A database of membrane spanning protein segments. Biol. Chem. Hoppe Seyler 347: 166.

    Google Scholar 

  • Hsu, L.C., Chiou, T.J., Chen, L. and Bush, D.R. 1993. Cloning a plant amino acid transporter by functional complementation of a yeast amino acid transport mutant. Proc. Natl. Acad. Sci. USA 90: 7441–7445.

    Google Scholar 

  • Jauniaux, J.C., Vandenbol, M., Vissers, S., Broman, K. and Grenson, M. 1987. Nitrogen catabolite regulation of proline permease in Saccharomyces cerevisiae. Cloning of the PUT4 gene and study of PUT4 RNA levels in wild-type and mutant strains. Eur. J. Biochem. 164: 601–606.

    Google Scholar 

  • Joshi, C.P. 1987. An inspection of the domain between putative TATAbox and translation start site in 79 plant genes. Nucl. Acids Res. 15: 6643–6653.

    Google Scholar 

  • Kay, R., Chan, A., Daly, M. and McPherson, J. 1987. Duplication of CaMV 35S promoter sequences creates a strong enhancer for plant genes. Science 236: 1299–1302.

    Google Scholar 

  • Kinraide, T.B. 1981. Electrical evidence for different mechanisms of uptake for basic, neutral and acidic amino acids in oat coleoptiles. Plant Physiol. 65: 1085–1089.

    Google Scholar 

  • Kinraide, T.B. and Etherton, B. 1980. Interamino acid inhibition of transport in higher plants. Plant Physiol. 68: 1327–1333.

    Google Scholar 

  • Kwart, M., Hirner, B., Hummel, S. and Frommer, W.B. 1993. Differential expression of two related amino acid transporters with differing substrate specificity in Arabidopsis thaliana. Plant J. 4: 993–1002.

    Google Scholar 

  • Li, Z.C. and Bush, D.R. 1990. 1pH-dependent amino acid transport into plasma membrane vesicles isolated from sugar beet leaves: I. Evidence for carrier-mediated, electrogenic flux through multiple transport systems. Plant Physiol. 94: 268–277.

    Google Scholar 

  • Li, Z.C. and Bush, D.R. 1991. 1pH-dependent amino acid transport into plasma membrane vesicles isolated from sugar beet (Beta vulgaris L.) leaves. II. Evidence for multiple aliphatic, neutral amino acid symports. Plant Physiol. 96: 1338–1344.

    Google Scholar 

  • Marvier, A.C., Neelam, A., Bick, J.A., Hall, J.L. and Williams, L.E. 1998. Cloning of a cDNA coding for an amino acid carrier from Ricinus communis (RcAAP1) by functional complementation in yeast: kinetic analysis, inhibitor sensitivity and substrate specificity. Biochim. Biophys. Acta 1373: 321–331.

    Google Scholar 

  • Mounoury, G., Delrot, S. and Bonnemain J. L. 1984. Energetics of threonine uptake by pod wall tissues of Vicia faba L. Planta 161: 178–185.

    Google Scholar 

  • Pate, J.S. 1973. Uptake, assimilation and transport of nitrogenous compounds by plants. Soil Biol. Biochem. 5: 109–119.

    Google Scholar 

  • Pate, J.S. 1980. Transport and partitioning of nitrogenous solutes. Annu. Rev. Plant Physiol. 31: 313–340.

    Google Scholar 

  • Pate, J.S. 1989. Origin, destination and fate of phloem solutes in relation to organ and whole plant functioning. In: D.A. Baker and J.A. Milburn (Eds.), Transport of Photoassimilates, Longman Scientific, London.

    Google Scholar 

  • Pate, J.S., Atkins, C.A., White, S.T., Rainbird, R.M. and Woo, K.C. 1980. Nitrogen nutrition and xylem transport of nitrogen in ureide-producing grain legumes. Plant Physiol. 65: 961–965.

    Google Scholar 

  • Rentsch, D., Hirner, B., Schmelzer, E. and Frommer, W.B. 1996. Salt stress-induced proline transporters and salt stress-repressed broad specificity amino acid permeases identified by suppression of a yeast amino acid permease-targeting mutant. Plant Cell 8: 1437–1446.

    Google Scholar 

  • Sambrook, J., Fritsch, E.F. and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.

    Google Scholar 

  • Sauer, N. and Stolz, J. 1994. SUC1 and SUC2: two sucrose transporters from Arabidopsis thaliana: expression and characterization in baker's yeast and identification of the histidine tagged protein. Plant J. 6: 67–77.

    Google Scholar 

  • Servaites, J.C., Schrader, L.E. and Jung, D.M. 1979. Energydependent loading of amino acids and sucrose into the phloem of soybean. Plant Physiol. 64: 546–550.

    Google Scholar 

  • Weston, K., Hall, J.L. and Williams, L.E. 1995. Characterization of amino-acid transport in Ricinus communis roots using isolated membrane vesicles. Planta 196: 166–173 (1995).

    Google Scholar 

  • Williams, L.E., Nelson, S.J. and Hall, J.L. 1992. Characterisation of solute proton cotransport in plasma membrane vesicles from Ricinus cotyledons, and a comparison with other tissues. Planta 186: 541–550.

    Google Scholar 

  • Winter, H., Lohaus, G. and Heldt, H.W. 1992. Phloem transport of amino acids and sucrose in correlation to the corresponding metabolite levels in barley leaves. Plant Physiol. 99: 996–1004.

    Google Scholar 

  • Wyse, R.E. and Komor, E. 1984. Mechanism of amino acid uptake by sugarcane suspension cells. Plant Physiol. 76: 865–870.

    Google Scholar 

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Authors and Affiliations

  1. Laboratoire de Physiologie et Biochimie Végétales, ERS CNRS 6099, B^atiment Botanique, UFR Sciences, Université de Poitiers, 40 avenue du Recteur Pineau, 86022, Poitiers Cédex, France

    Florence Montamat, Laurence Maurousset & Serge Delrot

  2. Plant Physiology, Zentrum für Molekularbiologie der Pflanzen (ZMBP), Universität Tübingen, Auf der Morgenstelle 1, 72076, Tübingen, Germany

    Mechthild Tegeder & Wolf Frommer

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  1. Florence Montamat
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  2. Laurence Maurousset
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  3. Mechthild Tegeder
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  4. Wolf Frommer
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  5. Serge Delrot
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Montamat, F., Maurousset, L., Tegeder, M. et al. Cloning and expression of amino acid transporters from broad bean. Plant Mol Biol 41, 259–268 (1999). https://doi.org/10.1023/A:1006321221368

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