Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1984 Jun;3(6):1389–1395. doi: 10.1002/j.1460-2075.1984.tb01982.x

Expression of the clustered mitochondrial tRNA genes in Saccharomyces cerevisiae: transcription and processing of transcripts.

C Palleschi, S Francisci, E Zennaro, L Frontali
PMCID: PMC557528  PMID: 6086323

Abstract

The transcripts of a cluster of eight tRNA genes localized in the Cap-oxiI region of the mitochondrial genome of Saccharomyces cerevisiae were investigated by hybridization of gene-specific probes on Northern blots of mitochondrial RNA and by S1 mapping of the 5' termini of the transcripts. Two rho- mutants that lack mature tRNA species and accumulate precursors have been used to detect transcripts that are not detectable in wild-type (w.t.) mitochondria. The results have shown the existence of polygenic transcripts carrying at least 5-7 tRNA sequences, both in w.t. and in rho- strains. The existence of several alternative processing pathways, which involve cleavage at the 3' and 5' ends of the tRNA sequences and in the long intergenic regions (possibly at GC clusters), is suggested. Cleavage at the 5' ends of tRNA sequences is defective in the mutant strains. The transcripts of the genes for tRNAThrACN and tRNACys (the tRNA genes immediately downstream from the 21S rRNA gene) have been analyzed; the possibility that these species represent primary transcripts is considered, and potential sites for initiation of transcription of the clustered tRNA genes are discussed.

Full text

PDF

Images in this article

1390Fig. 2.
on p.1390
1390Fig. 3.
on p.1390
1391Fig. 4.
on p.1391
1392Fig. 6.
on p.1392
1393Fig. 7.
on p.1393

Selected References

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

  1. Alwine J. C., Kemp D. J., Stark G. R. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5350–5354. doi: 10.1073/pnas.74.12.5350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baldacci G., Bernardi G. Replication origins are associated with transcription initiation sequences in the mitochondrial genome of yeast. EMBO J. 1982;1(8):987–994. doi: 10.1002/j.1460-2075.1982.tb01282.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  4. Berlani R. E., Bonitz S. G., Coruzzi G., Nobrega M., Tzagoloff A. Transfer RNA genes in the cap-oxil region of yeast mitochondrial DNA. Nucleic Acids Res. 1980 Nov 11;8(21):5017–5030. doi: 10.1093/nar/8.21.5017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Berlani R. E., Pentella C., Macino G., Tzagoloff A. Assembly of the mitochondrial membrane system: isolation of mitochondrial transfer ribonucleic acid mutants and characterization of transfer ribonucleic acid genes of Saccharomyces cerevisiae. J Bacteriol. 1980 Mar;141(3):1086–1097. doi: 10.1128/jb.141.3.1086-1097.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bonitz S. G., Tzagoloff A. Assembly of the mitochondrial membrane system. Sequences of yeast mitochondrial tRNA genes. J Biol Chem. 1980 Oct 10;255(19):9075–9081. [PubMed] [Google Scholar]
  7. Christianson T., Edwards J. C., Mueller D. M., Rabinowitz M. Identification of a single transcriptional initiation site for the glutamic tRNA and COB genes in yeast mitochondria. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5564–5568. doi: 10.1073/pnas.80.18.5564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Christianson T., Rabinowitz M. Identification of multiple transcriptional initiation sites on the yeast mitochondrial genome by in vitro capping with guanylyltransferase. J Biol Chem. 1983 Nov 25;258(22):14025–14033. [PubMed] [Google Scholar]
  9. Ciliberto G., Castagnoli L., Melton D. A., Cortese R. Promoter of a eukaryotic tRNAPro gene is composed of three noncontiguous regions. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1195–1199. doi: 10.1073/pnas.79.4.1195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Edwards J. C., Levens D., Rabinowitz M. Analysis of transcriptional initiation of yeast mitochondrial DNA in a homologous in vitro transcription system. Cell. 1982 Dec;31(2 Pt 1):337–346. doi: 10.1016/0092-8674(82)90127-1. [DOI] [PubMed] [Google Scholar]
  11. Frontali L., Palleschi C., Francisci S. Transcripts of mitochondrial tRNA genes in Saccharomyces cerevisiae. Nucleic Acids Res. 1982 Nov 25;10(22):7283–7293. doi: 10.1093/nar/10.22.7283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Galli G., Hofstetter H., Birnstiel M. L. Two conserved sequence blocks within eukaryotic tRNA genes are major promoter elements. Nature. 1981 Dec 17;294(5842):626–631. doi: 10.1038/294626a0. [DOI] [PubMed] [Google Scholar]
  13. Heckman J. E., Yin S., Alzner-DeWeerd B., RajBhandary U. L. Mapping and cloning of Neurospora crassa mitochondrial transfer RNA genes. J Biol Chem. 1979 Dec 25;254(24):12694–12700. [PubMed] [Google Scholar]
  14. Kole R., Baer M. F., Stark B. C., Altman S. E. coli RNAase P has a required RNA component. Cell. 1980 Apr;19(4):881–887. doi: 10.1016/0092-8674(80)90079-3. [DOI] [PubMed] [Google Scholar]
  15. Köchel H. G., Lazarus C. M., Basak N., Küntzel H. Mitochondrial tRNA gene clusters in Aspergillus nidulans: organization and nucleotide sequence. Cell. 1981 Feb;23(2):625–633. doi: 10.1016/0092-8674(81)90158-6. [DOI] [PubMed] [Google Scholar]
  16. Levens D., Ticho B., Ackerman E., Rabinowitz M. Transcriptional initiation and 5' termini of yeast mitochondrial RNA. J Biol Chem. 1981 May 25;256(10):5226–5232. [PubMed] [Google Scholar]
  17. Locker J. Analytical and preparative electrophoresis of RNA in agarose-urea. Anal Biochem. 1979 Oct 1;98(2):358–367. doi: 10.1016/0003-2697(79)90154-4. [DOI] [PubMed] [Google Scholar]
  18. Locker J., Rabinowitz M. Transcription in yeast mitochondria: analysis of the 21 S rRNA region and its transcripts. Plasmid. 1981 Nov;6(3):302–314. doi: 10.1016/0147-619x(81)90038-x. [DOI] [PubMed] [Google Scholar]
  19. Martin N. C., Underbrink-Lyon K. A mitochondrial locus is necessary for the synthesis of mitochondrial tRNA in the yeast Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4743–4747. doi: 10.1073/pnas.78.8.4743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  21. Miller D. L., Martin N. C. Characterization of the yeast mitochondrial locus necessary for tRNA biosynthesis: DNA sequence analysis and identification of a new transcript. Cell. 1983 Oct;34(3):911–917. doi: 10.1016/0092-8674(83)90548-2. [DOI] [PubMed] [Google Scholar]
  22. Montoya J., Christianson T., Levens D., Rabinowitz M., Attardi G. Identification of initiation sites for heavy-strand and light-strand transcription in human mitochondrial DNA. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7195–7199. doi: 10.1073/pnas.79.23.7195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Montoya J., Ojala D., Attardi G. Distinctive features of the 5'-terminal sequences of the human mitochondrial mRNAs. Nature. 1981 Apr 9;290(5806):465–470. doi: 10.1038/290465a0. [DOI] [PubMed] [Google Scholar]
  24. Morimoto R., Locker J., Synenki R. M., Rabinowitz M. Transcription, processing, and mapping of mitochondrial RNA from grande and petite yeast. J Biol Chem. 1979 Dec 25;254(24):12461–12470. [PubMed] [Google Scholar]
  25. Osinga K. A., De Haan M., Christianson T., Tabak H. F. A nonanucleotide sequence involved in promotion of ribosomal RNA synthesis and RNA priming of DNA replication in yeast mitochondria. Nucleic Acids Res. 1982 Dec 20;10(24):7993–8006. doi: 10.1093/nar/10.24.7993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Osinga K. A., Tabak H. F. Initiation of transcription of genes for mitochondrial ribosomal RNA in yeast: comparison of the nucleotide sequence around the 5'-ends of both genes reveals a homologous stretch of 17 nucleotides. Nucleic Acids Res. 1982 Jun 25;10(12):3617–3626. doi: 10.1093/nar/10.12.3617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Prunell A., Kopecka H., Strauss F., Bernardi G. The mitochondrial genome of wild-type yeast cells. V. Genome evolution. J Mol Biol. 1977 Feb 15;110(1):17–47. doi: 10.1016/s0022-2836(77)80096-x. [DOI] [PubMed] [Google Scholar]
  28. Sharp S., DeFranco D., Dingermann T., Farrell P., Söll D. Internal control regions for transcription of eukaryotic tRNA genes. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6657–6661. doi: 10.1073/pnas.78.11.6657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stark B. C., Kole R., Bowman E. J., Altman S. Ribonuclease P: an enzyme with an essential RNA component. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3717–3721. doi: 10.1073/pnas.75.8.3717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Underbrink-Lyon K., Miller D. L., Ross N. A., Fukuhara H., Martin N. C. Characterization of a yeast mitochondrial locus necessary for tRNA biosynthesis. Deletion mapping and restriction mapping studies. Mol Gen Genet. 1983;191(3):512–518. doi: 10.1007/BF00425771. [DOI] [PubMed] [Google Scholar]
  31. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Weaver R. F., Weissmann C. Mapping of RNA by a modification of the Berk-Sharp procedure: the 5' termini of 15 S beta-globin mRNA precursor and mature 10 s beta-globin mRNA have identical map coordinates. Nucleic Acids Res. 1979 Nov 10;7(5):1175–1193. doi: 10.1093/nar/7.5.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES