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Published on October 15, 2008; 10.1104/pp.108.130310

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Received September 25, 2008
Accepted October 10, 2008

Identification and characterisation of ADNT1, a novel mitochondrial adenine nucleotide transporter from Arabidopsis thaliana

Luigi Palmieri , Antonella Santoro , Fernando Carrari , Emanuela Blanco , Adriano Nunes-Nesi , Roberto Arrigoni , Francesco Genchi , Alisdair R. Fernie *, and Ferdinando Palmieri

Department of Pharmaco-Biology, Laboratory of Biochemistry and Molecular Biology, University of Bari, Via E. Orabona 4, 70125 Bari, Italy. CNR Institute of Biomembranes and Bioenergetics, Via E. Orabona 4, 70125 Bari, Italy. Department Willmitzer, Max-Planck-Institut fur Molekulare Pflanzenphysiologie, 14476 Potsdam-Golm, Germany

* Corresponding author; email: fernie{at}mpimp-golm.mpg.de.

Despite the fundamental importance and high level of compartmentation of mitochondrial nucleotide metabolism in plants, our knowledge concerning transport of nucleotides across intracellular membranes remains far from complete. Study of a previously uncharacterised gene (At4g01100) revealed it to be a novel adenine nucleotide transporter, designated as ADNT1, belonging to the mitochondrial carrier family. The ADNT1 gene shows broad expression at the organ level. Green fluorescent protein-based cell biological analysis demonstrated targeting of ADNT1 to mitochondria. Whilst analysis of the expression of GUS fusion proteins suggested that it was expressed across a broad range of tissue type it was most highly expressed in root tips. Direct transport assays with recombinant and reconstituted ADNT1 were utilized to demonstrate that this protein displays a relatively narrow substrate specificity largely confined to adenylates and their closest analogs. ATP uptake was markedly inhibited by the presence of other adenylates and general inhibitors of mitochondrial transport but not by bongkrekate or carboxyatractyloside, inhibitors of the previously characterised ADP/ATP carrier. Furthermore, the kinetics are substantially different from those of this carrier, with ADNT1 preferring AMP to ADP. Finally, isolation and characterisation of a T-DNA insertional knock-out mutant of ADNT1, alongside complementation and antisense approaches, demonstrated that although deficiency of this transporter did not seem to greatly alter photosynthetic metabolism it did result in reduced root growth and respiration. These findings are discussed in the context of a potential function for ADNT1 in the provision of the energy required to support growth in heterotrophic plant tissues.




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F. Palmieri, B. Rieder, A. Ventrella, E. Blanco, P. T. Do, A. Nunes-Nesi, A. U. Trauth, G. Fiermonte, J. Tjaden, G. Agrimi, et al.
Molecular Identification and Functional Characterization of Arabidopsis thaliana Mitochondrial and Chloroplastic NAD+ Carrier Proteins
J. Biol. Chem., November 6, 2009; 284(45): 31249 - 31259.
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