Molecular characterization and physiological function of plant nucleoside transporters

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Nucleoside transport across human, animal, fungi and protists has already been intensively characterised in the past. We have now identified the first plant nucleoside transporter ENT1 from Arabidopsis thaliana on a functional level. In the A. thaliana genome 8 different genes encode putative nucleoside transporters. Our aim is to determine their biochemical properties and analyse the corresponding physiological function. To do this the cDNA's will be cloned and expressed in heterologous systems (e.g. bakers yeast or xenopus oocytes) to determine the transport characteristics. Using GFP fusion proteins, which will be transiently expressed in protoplasts, we want to analyse the localisation of ENT isoforms within the cell. The activity of the various ENT promoters in different tissues, and during plant development will be analysed by generating transgenic plants with promoter-GUS fusions. Finally, the availability of knock out plants for ENT transporters for the first time provides insight in corresponding changes in phenotypes, development, and differentially expressed proteins.