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. 1980 Mar;77(3):1417–1421. doi: 10.1073/pnas.77.3.1417

Processing of precursors of 21S ribosomal RNA from yeast mitochondria.

S Merten, R M Synenki, J Locker, T Christianson, M Rabinowitz
PMCID: PMC348506  PMID: 6990410

Abstract

The transcription and processing of mitochondrial 21S rRNA in a petite strain of Saccharomyces cerevisiae has been examined by electron microscopic analysis of R-loop hybrids and by hybridization of labeled mitochondrial DNA probes to RNA transferred to diazobenzyloxymethyl paper. We have shown the presence of a large [5.1- to 5.4-kilobase (kb)] transcript that appears to be a precursor of mitochondrial 21S rRNA. This transcript contains sequences homologous to those of the mature 21S rRNA, to the intervening sequence present in the gene, and to additional sequences at the 3' end of the molecule. Our data suggest that this precursor of 21S rRNA is processed in two steps. The intron sequence is usually excised first, followed by removal of the extra 3' sequences. In some cases, however, the 3' extension is first removed and the intron sequence is then excised. Both pathways appear to lead to formation of the 3.1-kb mature 21S rRNA and a stable 1.2-kb intron transcript. Similar results were obtained with grande MH41-7B mitochondrial RNA by RNA transfer hybridization. We have also observed a number of additional transcripts that may be normal processing intermediates or may result from faulty cleavage-ligation during excision of the intervening sequence.

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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. Berg P. E., Lewin A., Christianson T., Rabinowitz M. Propagation of restriction fragments from the mitochondrial DNA of Saccharomyces cerevisiae in E. coli by means of plasmid vectors. Nucleic Acids Res. 1979;6(6):2133–2150. doi: 10.1093/nar/6.6.2133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bolotin-Fukuhara M., Fay G., Fukuhara H. Temperature-sensitive respiratory-deficient mitochondrial mutations: isolation and genetic mapping. Mol Gen Genet. 1977 Apr 29;152(3):295–305. doi: 10.1007/BF00693083. [DOI] [PubMed] [Google Scholar]
  4. Borst P., Grivell L. A. The mitochondrial genome of yeast. Cell. 1978 Nov;15(3):705–723. doi: 10.1016/0092-8674(78)90257-x. [DOI] [PubMed] [Google Scholar]
  5. Bos J. L., Heyting C., Borst P., Arnberg A. C., Van Bruggen E. F. An insert in the single gene for the large ribosomal RNA in yeast mitochondrial DNA. Nature. 1978 Sep 28;275(5678):336–338. doi: 10.1038/275336a0. [DOI] [PubMed] [Google Scholar]
  6. Casey J., Cohen M., Rabinowitz M., Fukuhara H., Getz G. S. Hybridization of mitochondrial transfer RNA's with mitochondrial and nuclear DNA of grande (wild type) yeast. J Mol Biol. 1972 Feb 14;63(3):431–440. doi: 10.1016/0022-2836(72)90438-x. [DOI] [PubMed] [Google Scholar]
  7. Faye G., Dennebouy N., Kujawa C., Jacq C. Inserted sequence in the mitochondrial 23S ribosomal RNA gene of the yeast Saccharomyces cerevisiae. Mol Gen Genet. 1979 Jan 5;168(1):101–109. doi: 10.1007/BF00267939. [DOI] [PubMed] [Google Scholar]
  8. Fukuhara H., Bolotin-Fukuhara M. Deletion mapping of mitochondrial transfer RNA genes in Saccharomyces cerevisiae by means of cytoplasmic petite mutants. Mol Gen Genet. 1976 Apr 23;145(1):7–17. doi: 10.1007/BF00331551. [DOI] [PubMed] [Google Scholar]
  9. Hensgens L. A., Grivell L. A., Borst P., Bos J. L. Nucleotide sequence of the mitochondrial structural gene for subunit 9 of yeast ATPase complex. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1663–1667. doi: 10.1073/pnas.76.4.1663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jakovcic S., Hendler F., Halbreich A., Rabinowitz M. Transcription of yeast mitochondrial deoxyribonucleic acid. Biochemistry. 1979 Jul 24;18(15):3200–3205. doi: 10.1021/bi00582a002. [DOI] [PubMed] [Google Scholar]
  11. Lewin A., Morimoto R., Rabinowitz M. Restriction enzyme analysis of mitochondrial DNAs of petite mutants of yeast: classification of petites, and deletion mapping of mitochondrial genes. Mol Gen Genet. 1978 Jul 25;163(3):257–275. doi: 10.1007/BF00271955. [DOI] [PubMed] [Google Scholar]
  12. Li M., Tzagoloff A. Assembly of the mitochondrial membrane system: sequences of yeast mitochondrial valine and an unusual threonine tRNA gene. Cell. 1979 Sep;18(1):47–53. doi: 10.1016/0092-8674(79)90352-0. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Locker J., Rabinowitz M., Getz G. S. Electron microscopic and renaturation kinetic analysis of mitochondrial DNA of cytoplasmic petite mutants of Saccharomyces cerevisiae. J Mol Biol. 1974 Sep 15;88(2):489–507. doi: 10.1016/0022-2836(74)90497-5. [DOI] [PubMed] [Google Scholar]
  15. Molloy P. L., Linnane A. W., Lukins H. B. Biogenesis of Mitochondria: Analysis of Deletion of Mitochondrial Antibiotic Resistance Markers in Petite Mutants of Saccharomyces cerevisiae. J Bacteriol. 1975 Apr;122(1):7–18. doi: 10.1128/jb.122.1.7-18.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Morimoto R., Lewin A., Rabinowitz M. Physical mapping and characterization of the mitochondrial DNA and RNA sequences from mit- mutants defective in cytochrome oxidase peptide 1 (OXI 3). Mol Gen Genet. 1979 Feb 16;170(1):1–9. doi: 10.1007/BF00268576. [DOI] [PubMed] [Google Scholar]
  17. Morimoto R., Lewin A., Rabinowitz M. Restriction cleavage map of mitochonrial DNA from the yeast Saccharomyces cerevisiae. Nucleic Acids Res. 1977 Jul;4(7):2331–2351. doi: 10.1093/nar/4.7.2331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  20. Schweyen R. J., Steyrer U., Kaudewitz F. Mapping of mitochondrial genes in Saccharomyces cerevisiae. Populations and pedigree analysis of retention or loss of four genetic markers in Rho-cells. Mol Gen Genet. 1976 Jul 23;146(2):117–132. doi: 10.1007/BF00268080. [DOI] [PubMed] [Google Scholar]
  21. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  22. Thomas M., White R. L., Davis R. W. Hybridization of RNA to double-stranded DNA: formation of R-loops. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2294–2298. doi: 10.1073/pnas.73.7.2294. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Van Ommen G. J., Groot G. S., Grivell L. A. Transcription maps of mtDNAs of two strains of saccharomyces: transcription of strain-specific insertions; Complex RNA maturation and splicing. Cell. 1979 Oct;18(2):511–523. doi: 10.1016/0092-8674(79)90068-0. [DOI] [PubMed] [Google Scholar]
  24. 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]

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