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Tomato EF-Tsmt, a functional mitochondrial translation elongation factor from higher plants

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Abstract

Ethylene-induced ripening in tomato (Lycopersicon esculentum) resulted in the accumulation of a transcript designated LeEF-Ts mt that encodes a protein with significant homology to bacterial Ts translational elongation factor (EF-Ts). Transient expression in tobacco and sunflower protoplasts of full-length and truncated LeEF-Ts mt -GFP fusion constructs and confocal microscopy observations clearly demonstrated the targeting of LeEF-Tsmt to mitochondria and not to chloroplasts and the requirement for a signal peptide for the proper sorting of the protein. Escherichia coli recombinant LeEF-Tsmt co-eluted from Ni-NTA resins with a protein corresponding to the molecular weight of the elongation factor EF-Tu of E. coli, indicating an interaction with bacterial EF-Tu. Increasing the GDP concentration in the extraction buffer reduced the amount of EF-Tu in the purified LeEF-Tsmt fraction. The purified LeEF-Tsmt stimulated the poly(U)-directed polymerization of phenylalanine 10-fold in the presence of EF-Tu. Furthermore, LeEF-Tsmt was capable of catalysing the nucleotide exchange reaction with E. coli EF-Tu. Altogether, these data demonstrate that LeEF-Tsmt encodes a functional mitochondrial EF-Ts. LeEF-Tsmt represents the first mitochondrial elongation factor to be isolated and functionally characterized in higher plants.

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Author notes

  1. Mohamed Benichou

    Present address: Faculty of Sciences, University of Cali Ayyad, BP 2390, Marrakech, Morocco

Authors and Affiliations

  1. UMR 990 INP/ENSAT-INRA, Avenue de l'Agrobiopole, BP 107, 31326, Castanet-Tolosan Cedex, France

    Mohamed Benichou, Barthélémy Tournier, Ana Chaves, Hicham Zegzouti, Corinne Delalande, Alain Latché, Mondher Bouzayen & Jean-Claude Pech

  2. Faculty of Sciences, University of Cali Ayyad, BP 2390, Marrakech, Morocco

    Alain Jauneau

  3. Department of Chemistry, University of North Carolina, Chapel Hill, NC, 27599-3290, USA

    Linda L. Spremulli

Authors
  1. Mohamed Benichou
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  2. Zhengguo Li
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  3. Barthélémy Tournier
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  10. Linda L. Spremulli
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  11. Jean-Claude Pech
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Correspondence to Jean-Claude Pech.

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Benichou, M., Li, Z., Tournier, B. et al. Tomato EF-Tsmt, a functional mitochondrial translation elongation factor from higher plants. Plant Mol Biol 53, 411–422 (2003). https://doi.org/10.1023/B:PLAN.0000006943.23747.7d

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