Summary
Yeast mitochondrial DNA codes for a complete set of tRNAs. Although most components necessary for the biosynthesis of mitochondrial tRNA are coded by nuclear genes, there is one genetic locus on mitochondrial DNA necessary for the synthesis of mitochondrial tRNAs other than the mitochondrial tRNA genes themselves. Characterization of mutants by deletion mapping and restriction enzyme mapping studies has provided a precise location of this yeast mitochondrial tRNA synthesis locus. Deletion mutants retaining various segments of mitochondrial DNA were examined for their ability to synthesize tRNAs from the genes they retain. A subset of these strains was also tested for the ability to provide the tRNA synthesis function in complementation tests with deletion mutants unable to synthesize mature mitochondrial tRNAs. By correlating the tRNA synthetic ability with the presence or absence of certain wild-type restriction fragments, we have confined the locus to within 780 base pairs of DNA located between the tRNA Metf gene and tRNAPro gene, at 29 units on the wild-type map. Heretofore, no genetic function or gene product had been localized in this area of the yeast mitochondrial genome.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Alwine JC, Kemp DJ, Stark GR (1977) Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci USA 74:5350–5354
Bos JL, Osingia KA, Vanderhorst G, Borst P (1979) Nucleotide sequences of the mitochondrial structural genes for cysteine tRNA and histidine tRNA of yeast. Nucl Acids Res 6:3255–3266
Casey J, Hsu HJ, Rabinowitz M, Getz G, Fukuhara H (1974) Transfer RNA genes in the mitochondrial DNA of cytoplasmic petite mutants of Saccharomyces cerevisiae. J Mol Biol 80:717–733
Debenham PG, Pongs O, Travers AA (1980) Formylmethionyl-tRNA alters RNA polymerase specificity. Proc Natl Acad Sci USA 77:870–874
Faye G, Bolotin-Fukuhara M, Fukuhara H (1976) Mitochondrial mutations that affect mitochondrial transfer ribonucleic acid in Saccharomyces cerevisiae. In: Bucher CT, Neupert W, Sebald W, Werner S (eds) The genetics and biogenesis of chloroplasts and mitochondria. North Holland Publishing Co, Amsterdam, pp 547–555
Frontali L, Palleschi C, Francisci S (1982) Transcripts of mitochondrial tRNA genes in Saccharomyces cerevisiae. Nucl Acids Res 10:7283–7293
Lewin A, Morimoto R, Rabinowitz M, Fukuhara H (1978) Restriction enzyme analysis of mitochondrial DNA's of petite mutants of yeast: Classification of petites, and deletion mapping of mitochondrial genes. Mol Gen Genet 163:257–275
Li M, Tzagoloff A (1979) Assembly of the mitochondrial membrane system: Sequences of yeast mitochondrial valine and an unusual threonine tRNA genes. Cell 18:47–53
Li M, Tzagoloff A, Underbrink-Lyon K, Martin NC (1982) Identification of the paromomycin-resistance mutation in the 15S rRNA gene of yeast mitochondria. J Biol Chem 257:5921–5928
Locker J (1979) Analytical preparative electrophoresis of RNA in agarose-urea gels. Biochemistry 98:358–367
Lopez IC, Farrelly F, Butow RA (1980) Trans action of the Var1 determinant region on yeast mitochondrial DNA. J Biol Chem 256:6496–6501
Martin N, Rabinowitz M, Fukuhara H (1976) Isoaccepting mitochondrial glutamyl tRNA species transcribed from different regions of the mitochondrial genome of Saccharomyces cerevisiae. J Mol Biol 101:285–296
Martin NC, Rabinowitz M, Fukuhara H (1977) Yeast mitochondrial DNA specifies tRNA for 19 amino acids. Deletion mapping of the tRNA genes. Biochemistry 16:4672–4677
Martin NC, Underbrink-Lyon K (1981) A mitochondrial locus is necessary for the synthesis of mitochondrial tRNA in the yeast Saccharomyces cerevisiae. Proc Natl Acad Sci USA 78:4743–4747
Miller DL, Martin NC (1981) Organization and expression of a tRNA gene cluster in Saccharomyces cerevisae mitochondrial DNA. Curr Genet 4:135–143
Miller DL, Martin NC, Pham HD, Donelson J (1979) Sequence analysis of two yeast mitochondrial DNA fragments containing the genes for tRNA SerUCR and tRNA PheUUY . J Biol Chem 254:11735–11740
Morimoto R, Locker J, Synenki RM, Rabinowitz M (1979) Transcription, processing, and mapping of mitochondrial RNA from grande and petite yeast. J Biol Chem 254:12461–12470
Morimoto R, Rabinowitz M (1979) Physical mapping of the yeast mitochondrial genome. Derivation of the fine structure and gene map of strain D273-10B and comparison with a strain (MH41-7B) differing in genome size. Mol Gen Genet 170:25–48
Newman D, Pham HD, Underbrink-Lyon K, Martin NC (1980) Characterization of tRNA genes in tRNA region II of yeast mitochondrial DNA. Nucl Acids Res 8:5007–5016
Nobrega FG, Tzagoloff A (1980) Assembly of the mitochondrial membrane system: Structure and location of the mitochondrial glutamic tRNA gene in Saccharomyces cerevisiae. FEBS Lett 113:52–54
Rogers D, Bussey H (1978) Fidelity of conjugation in Saccharomyces cerevisiae. Mol Gen Genet 162:173–182
Southern EM (1975) Transfer of DNA fragments to Millipore filters after restriction endonuclease cleavage followed by agarose gel electrophoresis. J Mol Biol 98:503–517
Stark B, Kole R, Bowman E, Altman S (1978) Ribonuclease P: An enzyme with an essential RNA component. Proc Natl Acad Sci USA 75:3717–3721
Stellwag EJ, Dahlberg A (1980) Electrophoretic transfer of DNA, RNA, and protein onto diazobenzyloxymethyl (DBM) paper. Nucl Acid Res 8:299–318
Vincent RD, Perlman PS, Strausberg RL, Butow RA (1980) Physical mapping of the genetic determinant on yeast mitochondrial DNA affecting the apparent size of the Var1 peptide. Curr Genet 2:27–38
Wesolowski M, Fukuhara H (1979) The genetic map of transfer RNA genes of yeast mitochondria: Correction and extension. Mol Gen Genet 170:261–275
Wesolowski M, Monnerot M, Fukuhara H (1980) MboI, ThaI, and Hinf I endonuclease cleavage maps of yeast mitochondrial DNA. Curr Genet 2:121–130
Author information
Authors and Affiliations
Additional information
Communicated by F. Kaudewitz
Rights and permissions
About this article
Cite this article
Underbrink-Lyon, K., Miller, D.L., Ross, N.A. et al. Characterization of a yeast mitochondrial locus necessary for tRNA biosynthesis. Molec. Gen. Genet. 191, 512–518 (1983). https://doi.org/10.1007/BF00425771
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00425771