Molecular and General Genetics MGG

, Volume 196, Issue 3, pp 465–472 | Cite as

The mitochondrial genome of the fission yeast Schizosaccharomyces pombe

2. Localization of genes by interspecific hybridization in strain ade7-50h- and cloning of the genome in small fragments
  • B. Franz Lang
  • Klaus Wolf
Article

Summary

  1. 1.

    A series of 18 small overlapping restriction fragments has been cloned, covering the complete mitochondrial genome of Schizosaccharomyces pombe.

     
  2. 2.

    By hybridizing mitochondrial gene probes from Saccharomyces cerevisiae and Neurospora crassa with restriction fragments of Schizosaccharomyces pombe mitochondrial DNA, the following homologous genes were localized on the mitochondrial genome of S. pombe: cob, cox1, cox2 and cox3, ATPase subunit 6 and 9 genes, the large rRNA gene and both types of open reading frames occurring in mitochondrial introns of various ascomycetes.

     
  3. 3.

    The region of the genome, hybridizing with cob exon probes is separated by an intervening sequence of about 2500 bp, which is homologous with the first two introns of the cox1 gene in Saccharomyces cerevisiae (class II introns according to Michel et al. 1982). Similarly, in the cox1 homologous region, which covers about 4000 bp, two regions were detected hybridizing with class I intron probes, suggesting the existence of two cox1 introns in Schizosaccharomyces pombe.

     
  4. 4.

    Hybridization with several specific exon probes with a determined order has revealed that cob, cox1, cox3 and the large rRNA gene are all transcribed from the same DNA strand.

     
  5. 5.

    The low intensities of hybridization signals suggest a large evolutionary distance between Schizosaccharomyces pombe and Saccharomyces cerevisiae or Neurospora crassa mitochondrial genes.

     
  6. 6.

    Considering the length of the mitochondrial DNA of Schizosaccharomyces pombe (about 19.4 kbp) and the expected length of the localized genes and intron sequences there is enough space left for encoding the expected set of tRNAs and the small rRNA gene.

     

The existence of leader-, trailer-, ori- and spacer sequences or further unassigned reading frames is then restricted to a total length of about 3000 bp only.

Abbreviations

MtDNA

mitochondrial DNA

cox1

cox2 and cox3 refer to the mitochondrial structural genes coding for three cytochrome oxidase subunits

ATPase

6 (oli2) and-9(oli1) to the two respective ATPase subunits in S. cerevisiae (Macino and Tzagoloff 1979, 1980)

cob

is the structural gene for apocytochrome b

var1

a mitochondrial gene of S. cerevisiae, coding for a protein subunit of the mitochondrial ribosomes (Hudspeth et al. 1982): The nomenclature of unassigned reading frames (urfs) refers to that introduced for HeLa mt DNA (Anderson et al. 1981) or S. cerevisiae (Macreadie et al. 1983; aap1 gene; Coruzzi et al. 1981: RF 1)

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References

  1. Anderson S, Bankier AT, Barrell BG, deBruijn HL, Coulson AR, Drouin J, Eperon IC, Nierlich DP, Roe BA, Sanger F, Schreier PH, Smith JH, Staden R, Young IG (1982) Comparison of the human and bovine mitochondrial genomes. In: Mitochondrial genes, Cold Spring Harbour Symposium. Cold Spring Harbour Laboratory, New York, pp 5–43Google Scholar
  2. Anziano PQ, Perlman PS, Lang BF, Wolf K (1983) The mitochondrial genome of the fission yeast Schizosaccharomyces pombe. I. Isolation and physical mapping of mitochondrial DNA. Curr Genet 7:273–284Google Scholar
  3. Beach D, Nurse P (1982) High frequency transformation of the fission yeast Schizosaccharomyces pombe. Nature 290:140–142Google Scholar
  4. Bechmann H, Haid A, Schweyen RJ, Mathews S, Kaudewitz F (1981) Expression of the “split gene” cob in yeast mt DNA. Translation of intervening sequences in mutant strains. J Biol Chem 256:3525–3531Google Scholar
  5. Benne R, Agostinelle M, de Vries BF, Van den Burg J, Klaver B, Borst P (1983) Gene expression and organization in trypanosome mitochondria. In: Schweyen RJ, Wolf K, Kaudewitz F (eds) Mitochondria 1983. Walter de Gruyter, Berlin, New York, pp 285–302Google Scholar
  6. Bernardi G (1975) Molecular Genetics of yeast mitochondria. In: Bandlow W, Schweyen RJ, Thomas DY, Wolf K, Kaudewitz F (eds) Genetics, biogenesis and bioenergetics of mitochondria. Walter de Gruyter, Berlin, New York, pp 69–84Google Scholar
  7. Bonitz SG, Coruzzi G, Thalenfeld E, Tzagoloff A, Macino G (1980) Assembly of the mitochondrial membrane system. Structure and nucleotide sequence of the gene coding for subunit 1 of yeast cytochrome oxidase. J Biol Chem 255: 11927–11941Google Scholar
  8. Burger G, Werner S (1983) Nucleotide sequence and transcript mapping of a mtDNA segment comprising CO 1, tRNA arg, and several unidentified reading frames in Neurospora crassa. In: Schweyen RJ, Wolf K, Kaudewitz F (eds) Mitochondria 1983. Walter de Gruyter, Berlin, New York, pp 331–342Google Scholar
  9. Burger G, Scriven C, Machleidt W, Werner S (1982) Subunit 1 of cytochrome oxidase from Neurospora crassa. Nucleotide sequence of the coding gene and partial amino acid sequence of the protein. The EMBO Journal 1:1385–1391Google Scholar
  10. Chang ACY, Cohen SN (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol 134:1141–1156Google Scholar
  11. Clark-Walker GD, Sriprakash KS (1981) Sequence rearrangements between mitochondrial DNAs of Torulopsis glabrata and Kloeckera africana identified by hybridization with six polypeptide encoding regions from Saccharomyces cerevisiae mitochondrial DNA. J Mol Biol 151:367–387Google Scholar
  12. Coruzzi G, Bonitz SG, Thalenfeld BE, Tzagoloff A (1981) Assembly of the mitochondrial membrane system. Analysis of the nucleotide sequence and transcripts in the oxil region of yeast mitochondrial DNA. J Biol Chem 256:12780–12787Google Scholar
  13. Davies RW, Waring RB, Brown TE (1983) Implications of fungal mitochondrial intron RNA structure for the mechanism for RNA splicing. In: Schweyen RJ, Wolf K, Kaudewitz F (eds) Mitochondria 1983. Walter de Gruyter, Berlin, New York, pp 179–189Google Scholar
  14. Dujon B (1983) Mitochondrial genes, mutants, maps — a review. In: Schweyen RJ, Wolf K, Kaudewitz F (eds) Mitochondria 1983. Walter de Gruyter, Berlin, New York, pp 1–24Google Scholar
  15. Fox TD (1979) Five TGA “stop” codons occur within the translated sequence of the yeast mitochondrial gene for cytochrome c oxidase subunit II Proc Natl Acad Sci USA:6534–6538Google Scholar
  16. Grivell LA (1983) Mitochondrial gene expression 1983. In: Schweyen RJ, Wolf K, Kaudewitz F (eds) Mitochondria 1983. Walter de Gruyter, Berlin, New York, pp 25–45Google Scholar
  17. Hensgens LAM, Grivell LA, Borst P, Bos JL (1979) Nucleotide sequence of the mitochondrial structural gene for subunit 9 of yeast ATPase complex. Proc Natl Acad Sci USA: 1663–1667Google Scholar
  18. Hensgens LAM, Bonen L, de Haan M, van der Horst G, Grivell LA (1983) Two intron sequences in yeast mitochondrial cox1 gene: Homology among URF-containing introns and straindependent variation in flanking exons. Cell 32:379–389Google Scholar
  19. Hudspeth MES, Ainley WM, Shumard DS, Butow RA, Grossman LI (1982) Location and structure of the var1 gene on yeast mitochondrial DNA: Nucleotide sequence of the 40.0 allele. Cell 30:617–626Google Scholar
  20. Macino G, Morelli G (1983) Cytochrome oxidase subunit 2 gene in Neurospora crassa mitochondria. J Biol Chem 258:13230–13235Google Scholar
  21. Macino G, Tzagoloff A (1979) Assembly of the mitochondrial membrane system. The DNA sequence of a mitochondrial ATPase gene in Saccharomyces cervisiae. J Biol Chem 254:4617–4623Google Scholar
  22. Macino G, Tzagoloff A (1980) Assembly of the mitochondrial membrane system. Sequence analysis of a yeast mitochondrial ATPase gene containing the oli-2 and oli-4 loci. Cell 20:507–517Google Scholar
  23. Macino G, Scazzocchio C, Waring RB, McPhail Berks M, Davies RW (1980) Conservation and rearrangement of mitochondrial structural gene sequences. Nature 288:404–406Google Scholar
  24. Macreadie IG, Novitzki CE, Maxwell RJ, John U, Ooi B, McMullen GL, Lukins HB, Linnane AW, Nagley P (1983) Biogenesis of mitochondria: the mitochondrial gene (aap1) coding for mitochondrial ATPase subunit 8 in Saccharomyces cerevisiae. Nucl Acids Res:4435–4451Google Scholar
  25. Michel F, Jacquier A, Dujon B (1982) Comparison of fungal mitochondrial introns reveals extensive homologies in RNA secondary structure. Biochimie 64:867–881Google Scholar
  26. Nobrega FG, Tzagoloff A (1980) Assembly of the mitochondrial membrane system. DNA sequence and organization of the cytochrome b gene in Saccharomyces cerevisiae. J Biol Chem 255:9828–9837Google Scholar
  27. O'Connor RM, McArthur CP, Clark-Walker GD (1975) Closed-Circular DNA from mitochondrial enriched fractions of four petite-negative yeasts. Eur J Biochem 53:137–144Google Scholar
  28. Rigby PWJ, Dieckmam M, Rhodes C, Berg P (1977) Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251Google Scholar
  29. Russel PR, Hall BD (1983) The primary structure of the alcohol dehydrogenase gene from the fission yeast Schizosaccharomyces pombe. J Biol Chem 258:143–149Google Scholar
  30. Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517Google Scholar
  31. Vincent RD, Perlman PS, Strausberg RL, Butow RA (1980) Physical mapping of genetic determinants on yeast mitochondrial DNA affecting the apparent size of the var1 — polypeptide. Curr Genet 2:27–38Google Scholar
  32. Viebrock A, Perz A, Sebald W (1982) The imported preprotein of the proteolipid subunit of the mitochondrial ATPsynthase from Neurospora crassa. Molecular cloning and sequencing of the mRNA. The EMBO Journal 1:565–571Google Scholar
  33. Wesolowski M, Algeri A, Fukuhara H (1981) Gene organization of the mitochondrial DNA of yeasts: Kluyveromyces lactis and Saccharomycopsis lipolytica. Curr Genet 3:157–162Google Scholar
  34. Wolf K, Dujon B, Slonimski PP (1973) Mitochondrial genetics. V. Multifactorial mitochondrial crosses involving a mutation conferring paromomycin — resistance in Saccharomyces cerevisiae. Mol Gen Genet 125:53–90Google Scholar
  35. Wolf K, Lang BF, del Giudice L, Anziano PQ, Perlman PS (1982) Schizosaccharomyces pombe: A short review of a short mitochondrial genome. In: Slonimski PP, Borst P, Attardi G (eds) Mitochondrial genes. Cold Spring Harbour Laboratory, New YorkGoogle Scholar
  36. Zimmer M, Lückemann G, Lang BF, Wolf K (1984) The mitochondrial genome of the fission yeast Schizosaccharomyces pombe. 3. Gene mapping in strain EF1 (CBS 356) and analysis of hybrids between the strains EF1 and ade7-50h. Mol Gen Genet 196:473–481Google Scholar

Copyright information

© Spriger-Verlag 1984

Authors and Affiliations

  • B. Franz Lang
    • 1
  • Klaus Wolf
    • 1
  1. 1.Institut für Genetik und Mikrobiologie der Universität MünchenMünchen 19Federal Republic of Germany

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