Abstract

Plants sequester intermediates of metabolic pathways into different cellular compartments, but the mechanisms by which these molecules are transported remain poorly understood. Monoterpene indole alkaloids, a class of specialized metabolites that includes the anticancer agent vincristine, antimalarial quinine and neurotoxin strychnine, are synthesized in several different cellular locations. However, the transporters that control the movement of these biosynthetic intermediates within cellular compartments have not been discovered. Here we present the discovery of a tonoplast localized nitrate/peptide family (NPF) transporter from Catharanthus roseus, CrNPF2.9, that exports strictosidine, the central intermediate of this pathway, into the cytosol from the vacuole. This discovery highlights the role that intracellular localization plays in specialized metabolism, and sets the stage for understanding and controlling the central branch point of this pharmacologically important group of compounds.

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Acknowledgements

This work was supported by grants from the European Research Council (311363), BBSRC (BB/J004561/1) (S.E.O.), Danish National Research Foundation (DNRF99 for D.X., V.N., M.B., H.H.N., B.A.H. and 10-082858 for F.G.-F.), the Innovation Fund Denmark (j.nr. 76-2014-3) (H.H.N.) and from the Région Centre, France (ABISAL grant; E.F., A.O., T.D.D.B., V.C.). R.M.E.P. was supported by a BBSRC Studentship.

Author information

Affiliations

  1. The John Innes Centre, Department of Biological Chemistry, Norwich Research Park, Norwich NR4 7UK, UK

    • Richard M. E. Payne
    • , Evangelos C. Tatsis
    •  & Sarah E. O'Connor
  2. DynaMo Centre, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, 40 Thorvaldsensvej, DK-1871 Frederiksberg C, Denmark

    • Deyang Xu
    • , Vlastimil Novak
    • , Meike Burow
    •  & Hussam Hassan Nour-Eldin
  3. Copenhagen Plant Science Centre, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, 1871 Frederiksberg C, Denmark

    • Deyang Xu
    • , Vlastimil Novak
    • , Meike Burow
    • , Carl-Erik Olsen
    • , Barbara Ann Halkier
    • , Fernando Geu-Flores
    •  & Hussam Hassan Nour-Eldin
  4. Université François-Rabelais de Tours, EA2106 Biomolécules et Biotechnologies Végétales, Département de Biologie et Physiologie Végétales, UFR Sciences et Techniques, Parc de Grandmont 37200 Tours, France

    • Emilien Foureau
    • , Marta Ines Soares Teto Carqueijeiro
    • , Audrey Oudin
    • , Thomas Dugé de Bernonville
    •  & Vincent Courdavault
  5. The John Innes Centre, Department of Computational and Systems Biology, Norwich Research Park, Norwich NR4 7UK, UK

    • D. Marc Jones
  6. The John Innes Centre, Department of Cell and Developmental Biology, Norwich Research Park, Norwich NR4 7UK, UK

    • Ali Pendle
  7. Section for Plant Biochemistry, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen, 1871 Frederiksberg C, Denmark

    • Fernando Geu-Flores

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Contributions

R.M.E.P. made the initial discovery of CrNPF2.9 and designed and carried out all silencing experiments; D.X. and H.H.N-E designed and carried out kinetic experiments; E.F., M.I.S.T.C., T.D.B. and V.C. designed and carried out localization experiments; F.G-F., S.E.O., H.H.N-E, V.C. and B.A.H. contributed to the conception and design of the experiments; R.M.E.P., D.X., E.F., M.I.S.T.C., A.O., T.D.B, V.N., M.B., C-E.O, D.M.J., E.C.T. and A.P. contributed to the acquisition of data; R.M.E.P., D.X., E.F., M.I.S.T.C., F.G-F., V.C., H.H.N-E. and S.E.O. contributed to data analysis and interpretation; R.M.E.P. and S.E.O. drafted the manuscript and all authors critically revised and approved the final version of the manuscript for publication.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Sarah E. O'Connor.

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