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MBC in Press, published online ahead of print April 2, 2004
Mol. Biol. Cell 10.1091/mbc.E03-11-0821

A more recent version of this article appeared on June 1, 2004
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Submitted on November 17, 2003
Revised on March 15, 2004
Accepted on March 19, 2004

The T-stem determines the cytosolic or mitochondrial localization of trypanosomal tRNAsMet

Anne Crausaz Esseiva1, Laurence Maréchal-Drouard2, Anne Cosset2, and André Schneider1*

1 Department of Biology/Zoology, University of Fribourg, Chemin du Musée 10, CH-1700 Fribourg, Switzerland
2 Institut de Biologie Moléculaire des Plantes du CNRS, UPR 2357, Université Louis Pasteur, 12 rue du Général Zimmer, 67084 Strasbourg Cedex, France

* Corresponding author. E-mail address: andre.schneider{at}unifr.ch.

The mitochondrion of T. brucei lacks tRNA genes. Organellar translation therefore depends on import of cytosolic, nucleus-encoded tRNAs. Except for the cytosol-specific initiator tRNAMet all trypanosomal tRNAs function in both the cytosol and the mitochondrion. The initiator tRNAMet is closely related to the imported elongator tRNAMet. Thus, the distinct localization of the two tRNAsMet must be specified by the 26 nucleotides which differ between the two molecules. Using transgenic T. brucei cell lines and subsequent cell fractionation we show that the T-stem is both required and sufficient to specify the localization of the tRNAsMet. Furthermore, it was shown that the tRNAMet T-stem localization determinants or also functional in the context of two other tRNAs. In vivo analysis of the modified nucleotides found in the initiator tRNAMet indicates that the T-stem localization determinants do not require modified nucleotides. In contrast, import of native tRNAsMet into isolated mitochondria suggests that nucleotide modifications might be involved in regulating the extent of import of elongator tRNAMet.




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