Plant Molecular Biology

, Volume 63, Issue 4, pp 505–517 | Cite as

Arabidopsis thaliana AtGpp1 and AtGpp2: two novel low molecular weight phosphatases involved in plant glycerol metabolism

  • José Antonio Caparrós-Martín
  • Sonja Reiland
  • Karl Köchert
  • Mari Cruz Cutanda
  • Francisco A. Culiáñez-Macià
Article

Abstract

We have isolated two Arabidopsis thaliana genes, AtGpp1 and AtGpp2, showing homology with the yeast low molecular weight phosphatases GPP1 and GPP2, which have a high specificity for dl-glycerol-3-phosphate, and moreover homology with DOG1 and DOG2 that dephosphorylate 2-deoxyglucose-6-phosphate. Using a comparative genomic approach, the corresponding genes were identified as conceptual translated haloacid dehalogenase-like hydrolase proteins. AtGpp1 (gi 18416631) and AtGpp2 (gi 18423981), encode proteins that share 95% identity, with a predicted Mw of 33 and 27 kDa and a pI of 7.8 and 5.6, respectively. Both isoforms have a high specificity for dl-glycerol-3-phosphate, pH optima at 7.0, and K m in the range of 3.5–5.2 mM. AtGpp1 and AtGpp2 are expressed throughout development in all plant organs, most strongly in siliqua, and expression is not affected by osmotic, ionic or oxidative stress. A putative chloroplast transit peptide cTP-containing sequence is appended to the AtGpp1 N-terminus while AtGpp2, devoid of this tail, is predicted to be in the extraplastidial cytosol; this compartmenting was further confirmed by subcellular fractionation. An immunohystochemical localization study, using anti-AtGpp2 antibodies, indicates that the AtGpp proteins are mainly restricted to the meristem of immature flower and vascular elements of the root, shoot, leave, siliqua and developing embryo. Considerable immunoreaction was observed in the cytoplasm as well as in plastid compartments of distinct cells types from different heterotrophic Arabidopsis tissues, and particularly localised within phloem companion cells. Transgenic Arabidopsis plants, with gain of AtGpp2 function, show altered phosphatase activity rates and improved tolerance to salt, osmotic and oxidative stress.

Keywords

Arabidopsis Glycerol metabolism Glycerol-3-phosphate Glycerol-3-phosphatases Transgenic plants Stress 

Notes

Acknowledgements

We acknowledge Professors Eduardo Primo-Yúfera (UPV, Valencia, Spain), Montserrat Pagès (CID-CSIC, Barcelona, Spain), Thomas Kupke (Lehrstuhl für Mikrobielle Genetik, Universität Tübingen, Germany), Francisco Montero and Olga Botella (Universidad de Castilla-La Mancha, Albacete, Spain) for their suggestions and warm support. We also thank the advice and provision of plasmid pJIT163 by Dr. Phil Mullineaux (John Innes Centre, Norwich, UK), pBin19 by Dr. Mike Bevan (John Innes Centre, Norwich, UK), and pSBETa by Dr. Florence Vignols and Yves Meyer (University of Perpignan, France); the antibodies production technical assistance by Dr. Concepción Cervera and Juan Carlos Moreno (UPV, Valencia, Spain); and the computer software help by Alexis González-Policarpo and Ramón Nogales-Rangel. This work was funded by the Research Project BIO2006-10138 from the MEC-FEDER of Spain.

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Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  • José Antonio Caparrós-Martín
    • 1
  • Sonja Reiland
    • 1
  • Karl Köchert
    • 1
  • Mari Cruz Cutanda
    • 1
  • Francisco A. Culiáñez-Macià
    • 1
  1. 1.Instituto de Biología Molecular y Celular de Plantas Primo-YúferaUniversidad Politécnica de Valencia-CSICValenciaSpain

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