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. 1994 Sep 11;22(18):3699–3705. doi: 10.1093/nar/22.18.3699

Nuclear-encoded mitochondrial tRNAs of Trypanosoma brucei have a modified cytidine in the anticodon loop.

A Schneider 1, K P McNally 1, N Agabian 1
PMCID: PMC308349  PMID: 7524027

Abstract

The mitochondrial genome of Trypanosoma brucei does not appear to encode any tRNA genes. Isolated organellar tRNAs hybridize to nuclear DNA, suggesting that they are synthesized in the nucleus and subsequently imported into the mitochondrion. Most imported tRNAs have cytosolic counterparts, showing identical mobility on two-dimensional polyacrylamide gels. We have compared three nuclear-encoded mitochondrial tRNAs (tRNA(Lys), tRNA(Leu), tRNA(Tyr)) with their cytosolic isoforms by direct enzymatic sequence analysis. Our findings indicate that the primary sequences of the mitochondrial and the corresponding cytosolic tRNAs are identical. However, we have identified a mitochondrion-specific nucleotide modification of each tRNA which is localized to a conserved cytidine residue at the penultimate position 5' of the anticodon. The modification present in mature mitochondrial tRNA(Tyr) was not found in a mutant tRNA(Tyr) defective in splicing in either cytosolic or mitochondrial fractions. The mutant tRNA(Tyr) has been expressed in transformed cells and its import into mitochondria has been demonstrated, suggesting that the modified cytidine residue is not required for import and therefore may be involved in adapting imported tRNAs to specific requirements of the mitochondrial translation machinery.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Braly P., Simpson L., Kretzer F. Isolation of kinetoplast-mitochondrial complexes from Leishmania tarentolae. J Protozool. 1974 Nov;21(5):782–790. doi: 10.1111/j.1550-7408.1974.tb03752.x. [DOI] [PubMed] [Google Scholar]
  2. Campbell D. A., Suyama Y., Simpson L. Genomic organisation of nuclear tRNAGly and tRNALeu genes in Trypanosoma brucei. Mol Biochem Parasitol. 1989 Dec;37(2):257–262. doi: 10.1016/0166-6851(89)90157-6. [DOI] [PubMed] [Google Scholar]
  3. Chen D. H., Shi X., Suyama Y. In vivo expression and mitochondrial import of normal and mutated tRNA(thr) in Leishmania. Mol Biochem Parasitol. 1994 Mar;64(1):121–133. doi: 10.1016/0166-6851(94)90140-6. [DOI] [PubMed] [Google Scholar]
  4. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  5. Dietrich A., Weil J. H., Maréchal-Drouard L. Nuclear-encoded transfer RNAs in plant mitochondria. Annu Rev Cell Biol. 1992;8:115–131. doi: 10.1146/annurev.cb.08.110192.000555. [DOI] [PubMed] [Google Scholar]
  6. Hancock K., Hajduk S. L. The mitochondrial tRNAs of Trypanosoma brucei are nuclear encoded. J Biol Chem. 1990 Nov 5;265(31):19208–19215. [PubMed] [Google Scholar]
  7. Hancock K., LeBlanc A. J., Donze D., Hajduk S. L. Identification of nuclear encoded precursor tRNAs within the mitochondrion of Trypanosoma brucei. J Biol Chem. 1992 Nov 25;267(33):23963–23971. [PubMed] [Google Scholar]
  8. Harris M. E., Moore D. R., Hajduk S. L. Addition of uridines to edited RNAs in trypanosome mitochondria occurs independently of transcription. J Biol Chem. 1990 Jul 5;265(19):11368–11376. [PubMed] [Google Scholar]
  9. Hausner T. P., Giglio L. M., Weiner A. M. Evidence for base-pairing between mammalian U2 and U6 small nuclear ribonucleoprotein particles. Genes Dev. 1990 Dec;4(12A):2146–2156. doi: 10.1101/gad.4.12a.2146. [DOI] [PubMed] [Google Scholar]
  10. Janke A., Päbo S. Editing of a tRNA anticodon in marsupial mitochondria changes its codon recognition. Nucleic Acids Res. 1993 Apr 11;21(7):1523–1525. doi: 10.1093/nar/21.7.1523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lonergan K. M., Gray M. W. Editing of transfer RNAs in Acanthamoeba castellanii mitochondria. Science. 1993 Feb 5;259(5096):812–816. doi: 10.1126/science.8430334. [DOI] [PubMed] [Google Scholar]
  12. Lustig F., Borén T., Claesson C., Simonsson C., Barciszewska M., Lagerkvist U. The nucleotide in position 32 of the tRNA anticodon loop determines ability of anticodon UCC to discriminate among glycine codons. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3343–3347. doi: 10.1073/pnas.90.8.3343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lye L. F., Chen D. H., Suyama Y. Selective import of nuclear-encoded tRNAs into mitochondria of the protozoan Leishmania tarentolae. Mol Biochem Parasitol. 1993 Apr;58(2):233–245. doi: 10.1016/0166-6851(93)90045-y. [DOI] [PubMed] [Google Scholar]
  14. Martin R. P., Schneller J. M., Stahl A. J., Dirheimer G. Import of nuclear deoxyribonucleic acid coded lysine-accepting transfer ribonucleic acid (anticodon C-U-U) into yeast mitochondria. Biochemistry. 1979 Oct 16;18(21):4600–4605. doi: 10.1021/bi00588a021. [DOI] [PubMed] [Google Scholar]
  15. Maréchal-Drouard L., Ramamonjisoa D., Cosset A., Weil J. H., Dietrich A. Editing corrects mispairing in the acceptor stem of bean and potato mitochondrial phenylalanine transfer RNAs. Nucleic Acids Res. 1993 Oct 25;21(21):4909–4914. doi: 10.1093/nar/21.21.4909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Maréchal-Drouard L., Weil J. H., Guillemaut P. Import of several tRNAs from the cytoplasm into the mitochondria in bean Phaseolus vulgaris. Nucleic Acids Res. 1988 Jun 10;16(11):4777–4788. doi: 10.1093/nar/16.11.4777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mottram J. C., Bell S. D., Nelson R. G., Barry J. D. tRNAs of Trypanosoma brucei. Unusual gene organization and mitochondrial importation. J Biol Chem. 1991 Sep 25;266(27):18313–18317. [PubMed] [Google Scholar]
  18. Noller H. F. tRNA-rRNA interactions and peptidyl transferase. FASEB J. 1993 Jan;7(1):87–89. doi: 10.1096/fasebj.7.1.8422979. [DOI] [PubMed] [Google Scholar]
  19. Schneider A. Import of RNA into mitochondria. Trends Cell Biol. 1994 Aug;4(8):282–286. doi: 10.1016/0962-8924(94)90218-6. [DOI] [PubMed] [Google Scholar]
  20. Schneider A., Martin J., Agabian N. A nuclear encoded tRNA of Trypanosoma brucei is imported into mitochondria. Mol Cell Biol. 1994 Apr;14(4):2317–2322. doi: 10.1128/mcb.14.4.2317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schneider A., McNally K. P., Agabian N. Splicing and 3'-processing of the tyrosine tRNA of Trypanosoma brucei. J Biol Chem. 1993 Oct 15;268(29):21868–21874. [PubMed] [Google Scholar]
  22. Simpson A. M., Suyama Y., Dewes H., Campbell D. A., Simpson L. Kinetoplastid mitochondria contain functional tRNAs which are encoded in nuclear DNA and also contain small minicircle and maxicircle transcripts of unknown function. Nucleic Acids Res. 1989 Jul 25;17(14):5427–5445. doi: 10.1093/nar/17.14.5427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Simpson L. RNA editing--a novel genetic phenomenon? Science. 1990 Oct 26;250(4980):512–513. doi: 10.1126/science.1700474. [DOI] [PubMed] [Google Scholar]
  24. Small I., Maréchal-Drouard L., Masson J., Pelletier G., Cosset A., Weil J. H., Dietrich A. 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? EMBO J. 1992 Apr;11(4):1291–1296. doi: 10.1002/j.1460-2075.1992.tb05172.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sprinzl M., Hartmann T., Meissner F., Moll J., Vorderwülbecke T. Compilation of tRNA sequences and sequences of tRNA genes. Nucleic Acids Res. 1987;15 (Suppl):r53–188. doi: 10.1093/nar/15.suppl.r53. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. de Zamaroczy M., Bernardi G. The primary structure of the mitochondrial genome of Saccharomyces cerevisiae--a review. Gene. 1986;47(2-3):155–177. doi: 10.1016/0378-1119(86)90060-0. [DOI] [PubMed] [Google Scholar]
  27. de la Cruz V. F., Lake J. A., Simpson A. M., Simpson L. A minimal ribosomal RNA: sequence and secondary structure of the 9S kinetoplast ribosomal RNA from Leishmania tarentolae. Proc Natl Acad Sci U S A. 1985 Mar;82(5):1401–1405. doi: 10.1073/pnas.82.5.1401. [DOI] [PMC free article] [PubMed] [Google Scholar]

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