In vivo import of a normal or mutagenized heterologous transfer RNA into the mitochondria of transgenic plants: towards novel ways of influencing mitochondrial gene expression?

Small, Ian; Maréchal-Drouard, Laurence; Masson, Jean; Pelletier, Georges; Cosset, Anne; Weil, Jacques-Henry; Dietrich, André

doi:10.1002/j.1460-2075.1992.tb05172.x

Cited by 107 publications

(69 citation statements)

References 43 publications

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“…The occurrence in mitochondria of nucleus-encoded RNA species has been previously reported, for tRNAs, in plants (59), Tetrahymena thermophila (53,54), Leishmania (1), Trypanosoma brucei (24), and S. cerevisiae (38), for MRP RNA, in mouse L cells (11) and mouse cardiomyocytes and myogenic cells (34), and, more recently, for 5S rRNA, in mammalian cells (36). In addition human immunodeficiency virus RNA has been detected in mitochondria of infected cells (60).…”

Section: Discussionmentioning

confidence: 62%

The RNase P Associated with HeLa Cell Mitochondria Contains an Essential RNA Component Identical in Sequence to That of the Nuclear RNase P

Puranam

Attardi

2001

Molecular and Cellular Biology

122

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The mitochondrion-associated RNase P activity (mtRNase P) was extensively purified from HeLa cells and shown to reside in particles with a sedimentation constant (ϳ17S) very similar to that of the nuclear enzyme (nuRNase P). Furthermore, mtRNase P, like nuRNase P, was found to process a mitochondrial tRNA Ser(UCN) precursor [ptRNA Ser(UCN) ] at the correct site. Treatment with micrococcal nuclease of highly purified mtRNase P confirmed earlier observations indicating the presence of an essential RNA component. Furthermore, electrophoretic analysis of 3-end-labeled nucleic acids extracted from the peak of glycerol gradientfractionated mtRNase P revealed the presence of a 340-nucleotide RNA component, and the full-length cDNA of this RNA was found to be identical in sequence to the H1 RNA of nuRNase P. The proportions of the cellular H1 RNA recovered in the mitochondrial fractions from HeLa cells purified by different treatments were quantified by Northern blots, corrected on the basis of the yield in the same fractions of four mitochondrial nucleic acid markers, and shown to be 2 orders of magnitude higher than the proportions of contaminating nuclear U2 and U3 RNAs. In particular, these experiments revealed that a small fraction of the cell H1 RNA (of the order of 0.1 to 0.5%), calculated to correspond to ϳ33 to ϳ175 intact molecules per cell, is intrinsically associated with mitochondria and can be removed only by treatments which destroy the integrity of the organelles. In the same experiments, the use of a probe specific for the RNA component of RNase MRP showed the presence in mitochondria of 6 to 15 molecules of this RNA per cell. The available evidence indicates that the levels of mtRNase P detected in HeLa cells should be fully adequate to satisfy the mitochondrial tRNA synthesis requirements of these cells.The unique mode of transcription of the mammalian mitochondrial DNA in the form of giant polycistronic molecules, containing tRNA sequences regularly interspersed between the individual rRNA and mRNA sequences and in most cases butt-joined to them (39,42,43), demands the existence of a complex RNA-processing apparatus. The tRNA sequences presumably function as signals for RNA-processing enzymes, which carry out the endonucleolytic cleavages that eventually lead to the formation of the mature rRNA, mRNA, and tRNA species (43). One of these enzymatic activities would be expected to cut precisely the polycistronic transcripts on the 5Ј side of each tRNA sequence and therefore to be analogous to the RNase P, an RNA-containing enzyme first identified in Escherichia coli (3), and subsequently found ubiquitously in prokaryotic and eukaryotic organisms (4). In previous work from this laboratory, an endoribonuclease which cleaves the precursor of the E. coli suppressor tRNA Tyr at the same site as E. coli RNase P, producing the mature 5Ј end of this tRNA, has been identified. The enzyme was partially purified from HeLa cell mitochondria (and is henceforth referred to as mtRNase P) (15). An analogous enzyme ...

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Section: Discussionmentioning

confidence: 62%

The RNase P Associated with HeLa Cell Mitochondria Contains an Essential RNA Component Identical in Sequence to That of the Nuclear RNase P

Puranam

Attardi

2001

Molecular and Cellular Biology

122

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“…Using oligonucleotide-directed mutagenesis (18), these fragments were fused downstream of a 178-nucleotide-long fragment containing the promoter sequence of the potato mitochondrial 18 S rRNA gene ((19) amplified by PCR from potato (Solanum tuberosum) mitochondrial DNA (Fig. 1).…”

Section: Methodsmentioning

confidence: 99%

Fate of a Larch Unedited tRNA Precursor Expressed in Potato Mitochondria

Placido

Gagliardi

Gallerani

et al. 2005

Journal of Biological Chemistry

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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.

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“…It is recognized that the RNA moieties of RNase MRP, which are known to process mitochondrial RNAs, are encoded by nuclear genomes of animals [13]. Recently, the import of tRNAs into mitochondria was demonstrated in vivo in S. cerevisiue [6] and in plants [26], as well as in vitro in S. cerevisiue [6]. In this case an energy source and cytoplasmic protein factor(s) are required for the tRNA transport, which is similar to the case of protein transport.…”

mentioning

confidence: 99%

Existence of nuclear‐encoded 5S‐rRNA in bovine mitochondria

et al. 1994

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A number of proteins functioning in mitochondria are synthesized in the cytoplasm and imported into the mitochondria via specific transport systems. In mammals, on the contrary, mitochondrial membranes have generally been considered to be impermeable to nucleic acids. However, here we show that an RNA with 120 nucleotides, the sequence of which is identical to that of the nuclear-encoded 5s RNA, exists in bovine mitochondria, although the mitochondrial genome encodes no 5s RNA gene. This RNA molecule was found to be retained in purified bovine mitochondria as well as in the mitoplasts, even after extensive treatment with an RNase, demonstrating that the 5s RNA is actually located inside the mitochondrial inner membrane. The 5s rRNA molecule was also shown to exist in mitochondria from rabbit and chicken.

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In vivo import of a normal or mutagenized heterologous transfer RNA into the mitochondria of transgenic plants: towards novel ways of influencing mitochondrial gene expression?

Cited by 107 publications

References 43 publications

The RNase P Associated with HeLa Cell Mitochondria Contains an Essential RNA Component Identical in Sequence to That of the Nuclear RNase P

The RNase P Associated with HeLa Cell Mitochondria Contains an Essential RNA Component Identical in Sequence to That of the Nuclear RNase P

Fate of a Larch Unedited tRNA Precursor Expressed in Potato Mitochondria

Existence of nuclear‐encoded 5S‐rRNA in bovine mitochondria

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