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J. Biol. Chem., Vol. 280, Issue 39, 33573-33579, September 30, 2005

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Fate of a Larch Unedited tRNA Precursor Expressed in Potato Mitochondria^*

Antonio Placido1, Dominique Gagliardi, Raffaele Gallerani, Jean-Michel Grienenberger, and Laurence Maréchal-Drouard2

From the Dipartimento di Biochimica e Biologia Molecolare, Universita' di Bari, CNR, Via Orabona 4, 70126 Bari, Italy and the Institut de Biologie Moléculaire des Plantes, UPR2357 CNRS, 12 rue du Général Zimmer, 67084 Strasbourg Cedex, France

In higher plant mitochondria, post-transcriptional C to U conversion known as editing mostly affects mRNAs. However, three tRNAs were also shown to be edited. Among them, three editing sites were identified in larch mitochondrial tRNA^His. We have previously shown that only the edited version can undergo maturation in vitro. In this paper, we introduced via direct DNA uptake the edited or unedited version of larch mitochondrial trnH into isolated potato mitochondria and expressed them under the control of potato mitochondrial 18 S rRNA promoter. As expected, the edited form of larch mitochondrial tRNA^His precursor was processed in the isolated organelles. By contrast, no mature tRNA^His was detected when using the unedited version of trnH. However, precursor molecules could be characterized by reverse transcription-PCR. These data demonstrate that the potato mitochondrial editing machinery is not able to recognize these "foreign" editing sites and confirm that these unedited tRNA precursor molecules are not correctly processed in organello. As a consequence, the fate of these RNA precursor molecules is likely to be degradation. Indeed, we detected by PCR two 3'-end truncated precursor RNAs. Interestingly, both RNA species exhibit poly(A) tails, a hallmark of degradation in plant mitochondria. Taken together, these data suggest that, in plant mitochondria, a defective unedited RNA precursor that cannot be processed to give a mature stable tRNA, is degraded through a polyadenylation-dependent pathway.

Received for publication, May 13, 2005 , and in revised form, July 27, 2005.

^* This work was supported in part by the Centre National de la Recherche Scientifique (CNRS, France) and by Dipartimento di Biochimica e Biologia Molecolare (Bari University, Italy). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Supported by a Marie Curie host fellowship.

² To whom correspondence should be addressed: Inst. de Biologie Moléculaire des Plantes, UPR2357 CNRS, 12 rue du Général Zimmer, 67084 Strasbourg Cedex, France. Tel.: 33-3-88-41-72-44; Fax: 33-3-88-61-44-42; E-mail: laurence.drouard{at}ibmp-ulp.u-strabg.fr.

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