DNA sequence polymorphisms in the genus Saccharomyces. I. Comparison of theHIS4 and ribosomal RNA genes in lager strains, ale strains and various species

  • Mogens Bohl Pedersen
Article

Abstract

The region of chromosome XII containing theRDN1 gene which encodes the cytosolic ribosomal RNA molecules and the region of chromosome III containing theHIS4 (histidine 4) gene were analysed in 30 lager yeast strains, 11 ale strains and 20 strains from a number of different species in the genus Saccharomyces.

With the aid of restriction endonuclease fragment patterns and cloned probes to theRDN1 gene of S. cerevisiae three forms of this gene were identified, two of them corresponding to the previously known forms I and II and a third one characterized by an additional HindIII site located in the 3′ spacer region. A more distantly related form of theRDN1 gene containing a single HindIII restriction site was found in Saccharomyces fermentati and one form without any HindIII site in a wild yeast contaminant.

With the help of the restriction endonuclease fragments derived from theHIS4 region seven genotypes can be recognized. They result from various combinations of three restriction endonuclease fragment patterns designated I, II and III, each pattern represents a chromosome.

All lager strains are homozygous for form II of the ribosomal RNA gene and heterozygous for patterns I and II of theHIS4 gene. An exception is one German brewing strain which is homozygous for pattern II.

With one exception the ale strains were homozygous for form II of theRDN1 gene and for pattern I of theHIS4 gene. One British strain contains form I of theRDN1 gene. Bakers yeast, S. diastaticus and S. italicus are homozygous for form I of theRDN1 gene and for pattern I of theHIS4 gene. In S. bayanus and S. pastorianus homozygosity for form III of theRDN1 gene was combined with heterozygosity for patterns II and III of theHIS4 gene. S. uvarum is homozygous for both form III of theRDN1 gene and pattern III of theHIS4 gene. Form III of theRDN1 gene and patterns I and II of theHIS4 gene were combined in a Chinese brewing strain and a strain designated as a type strain of S. carlsbergensis.

The nucleotide sequence polymorphisms are useful markers for strain characterization in addition to the generally used fermentation properties.

Keywords

Brewers yeasts α-galactosidase sporulation restriction endonuclease patterns molecular hybridization S. bayanus S. carlsbergensis S. cerevisiae S. diastaticus S. ellipsoideus S. fermentati S. italicus S. pastorianus 

Abbreviations

bp

basepairs

kb

kilobase

SSC

0.15m-NaCl, 15mm-Na citrate

Tris

tris-(hydroxymethyl)-amino methane

References

  1. 1.
    Bicknell, J.N. &H.C. Douglas: Nucleic acid homologies among species of Saccharomyces. J. Bacteriol. 101, 505–512 (1970)PubMedGoogle Scholar
  2. 2.
    Buckholz, R.G. &B.G. Adams: Induction and genetics of two α-galactosidase activities in Saccharomyces cerevisiae. Mol. Gen. Genet. 182, 77–81 (1981)PubMedCrossRefGoogle Scholar
  3. 3.
    Cameron, J.R., P. Philippsen &R.W. Davis: Analysis of chromosomal integration and deletions of yeast plasmids. Nucl. Acids Res. 4, 1429–1448 (1977)PubMedCrossRefGoogle Scholar
  4. 4.
    Carlson M. &D. Botstein: Organization of the SUC gene family in Saccharomyces. Mol. Cell. Biol. 3, 351–359 (1983)PubMedGoogle Scholar
  5. 5.
    Cramer, J.H., F.W. Farrelly &R.H. Rownd: Restriction endonuclease analysis of ribosomal DNA from Saccharomyces cerevisiae. Mol. Gen. Genet. 148, 233–241 (1976)PubMedCrossRefGoogle Scholar
  6. 6.
    Delihas, N. &J. Andersen: Generalized structures of the 5S ribosomal RNAs. Nucl. Acids Res. 10, 7323–7344 (1982)PubMedCrossRefGoogle Scholar
  7. 7.
    Denhardt, D.T.: A membrane-filter technique for the detection of complementary DNA. Biochem. Biophys. Res. Comm. 23, 641–646 (1966)PubMedCrossRefGoogle Scholar
  8. 8.
    Donhauser, S., H. Ritter & J. Schmitt: Genetische Variabilität der Alkoholdehydrogenasen bei Saccharomyces. Proc. 18th Europ. Brewery Conv. Congr. Copenhagen. 187–196 (1981)Google Scholar
  9. 9.
    Erdmann, V.A., E. Huysmans, A. Vanderberghe &R. De Wachter: Collection of published 5S and 5.8S ribosomal sequences. Nucl. Acids Res. 11, r105-r123 (1983)PubMedCrossRefGoogle Scholar
  10. 10.
    Gjermansen, C. &P. Sigsgaard: Construction of a hybrid brewing strain of Saccharomyces carlsbergensis by mating of meiotic segregants. Carlsberg Res. Commun. 46, 1–11 (1981)CrossRefGoogle Scholar
  11. 11.
    Holmberg, S., J.G.L. Petersen, T. Nilsson-Tillgren &M.C. Kielland-Brandt: Molecular characterization of a Saccharomyces plasmid containing theHIS4 gene. Carlsberg Res. Commun. 44, 269–282 (1979)CrossRefGoogle Scholar
  12. 12.
    Holmberg, S.: Genetic differences between Saccharomyces carlsbergensis and S. cerevisiae II. Restriction endonuclease analysis of genes in chromosome III. Carlsberg Res. Commun. 47, 233–244 (1982)CrossRefGoogle Scholar
  13. 13.
    Holmes, D.S. &M. Quigley: A rapid boiling method for the preparation of bacterial plasmids. Anal. Biochem. 114, 193–197 (1981)PubMedCrossRefGoogle Scholar
  14. 14.
    Jeffreys, A.J. &R.A. Flavell: A physical map of the DNA regions flanking the rabbit β-globin gene. Cell 12, 429–439 (1977)PubMedCrossRefGoogle Scholar
  15. 15.
    Johannsen, E. &J.P. van der Walt: Interfertility as basis for the delimitation of Kluyveromyces marxianus. Arch. Microbiol. 118, 45–48 (1978)CrossRefGoogle Scholar
  16. 16.
    Maniatis T., E.F. Fritsch & J. Sambrook: Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory (1982)Google Scholar
  17. 17.
    Nilsson-Tillgren, T., C. Gjermansen, M.C. Kielland-Brandt, J.G.L. Petersen &S. Holmberg: Genetic differences between Saccharomyces carlsbergensis and S. cerevisiae. Analysis of chromosome III by single chromosome transfer. Carlsberg Res. Commun. 46, 65–76 (1981)CrossRefGoogle Scholar
  18. 18.
    Peacock, A.C. &C.W. Dingman: Molecular weight estimation and separation of ribonucleic acid by electrophoresis in agarose-acrylamide composite gels. Biochemistry 7, 668–674 (1968)PubMedCrossRefGoogle Scholar
  19. 19.
    Pedersen, M.B.: Fingerprinting of DNA in the identification of yeast. Proc. 19th Europ. Brewery Conv. Congr. London, pp. 457–463 (1983)Google Scholar
  20. 20.
    Petes, T.D., L.M. Hereford &K.G. Skryabin: Characterization of two types of yeast ribosomal DNA genes. J. Bacteriol. 134, 295–305 (1978)PubMedGoogle Scholar
  21. 21.
    Petes, T.D., S. Smolik-Utlaut &T. Zamb: Genetic analysis of the repeating ribosomal DNA genes of yeast. In: Molecular Genetics in Yeast. Alfred Benzon Symposium 16. Eds: D. von Wettstein, J. Friis, M. Kielland-Brandt & A. Stenderup. Munksgaard. Copenhagen, pp. 137–151 (1981)Google Scholar
  22. 22.
    Petes, T.D.: Meiotic mapping of yeast ribosomal deoxyribonucleic acid on chromosome XII. J. Bacteriol. 138, 185–192 (1979)PubMedGoogle Scholar
  23. 23.
    Philippsen, P., R.A. Kramer &R.W. Davis: Cloning of the yeast ribosomal DNA repeat unit in SstI and HindIII lambda vectors using genetic and physical size selection. J. Mol. Biol. 123, 371–386 (1978)PubMedCrossRefGoogle Scholar
  24. 24.
    Philippsen, P., M. Thomas, R.A. Kramer &R.W. Davis: Unique arrangement of coding sequences for 5S, 5.8S, 18S, 25S ribosomal RNA in Saccharomyces cerevisiae as determined by R-loop and hybridization analysis. J. Mol. Biol. 123, 287–404 (1978)CrossRefGoogle Scholar
  25. 25.
    Price, C.W., G.B. Fuson &H.J. Phaff: Genome comparison in yeast systematics: Species within the genera Schwanniomyces, Saccharomyces, Debaryomyces and Picia. Microbiol. Rev. 42, 161–193 (1978)PubMedGoogle Scholar
  26. 26.
    Rigby, P.W.J., M. Dieckmann, C. Rhodes &P. Berg: Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J. Mol. Biol. 113, 237–251 (1977)PubMedCrossRefGoogle Scholar
  27. 27.
    Rosenthal, A.L. &A. Saifer: Continuous monitoring of fluorogenic substrates I. Kinetic analysis of N-acetyl-β-D-hexosaminidases. Anal. Biochem. 55, 85–92 (1973)PubMedCrossRefGoogle Scholar
  28. 28.
    Rosini, G., F. Federici, A.E. Vaughan &A. Martini: Systematics of the species of the genus Saccharomyces associated with the fermentation industry. European J. Appl. Microbiol. Biotechnol. 15, 188–193 (1982)CrossRefGoogle Scholar
  29. 29.
    Schweitzer, E., C. MacKechnie &H.O. Halvorson: The redundancy of ribosomal and transfer RNA genes in Saccharomyces cerevisiae. J. Mol. Biol. 40, 261–277 (1969)CrossRefGoogle Scholar
  30. 30.
    Sherman, F., G.R. Fink &J.B. Hicks: Methods in Yeast Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y. 11724. pp 61–64 (1979)Google Scholar
  31. 31.
    Southern, E.M.: Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98, 504–517 (1975)CrossRefGoogle Scholar
  32. 32.
    Torczynski, R., A.P. Bollon &M. Fuke: The complete nucleotide sequence of the rat 18S ribosomal RNA gene and comparison with the respective yeast and frog genes. Nucl. Acids Res. 11, 4879–4890 (1983)PubMedCrossRefGoogle Scholar
  33. 33.
    van der Walt, J.P.: Genus Saccharomyces. In: The Yeasts, a Taxonomic Study. J. Lodder, ed. North-Holland Publishing Company. Amsterdam London pp. 555–718 (1970)Google Scholar
  34. 34.
    Verbeet, M.Ph., J. Klootwijk, H. van Heerikshuisen, R. Fontijn, E. Vreugdenhil &R.J. Planta: Molecular cloning of the rDNA of Saccharomyces rosei and comparison of its transcription initiation region with that of Saccharomyces carlsbergensis. Gene 23, 53–63 (1983)PubMedCrossRefGoogle Scholar
  35. 35.
    Wettstein, D. von: Emil Christian Hansen Centennial Lecture. From pure culture to genetic engineering of brewers yeast. Proc. 19th Europ. Brewery Conv. Congr. London, pp. 97–119 (1983)Google Scholar
  36. 36.
    Winge, Ø. &O. Laustsen: On 14 new yeast types, produced by hybridization. Compt. Rend. Lab. Carlsberg, Ser. Physiol., 22, 337–352 (1939)Google Scholar
  37. 37.
    Yamazak, M., S. Goto &K. Komagata: Comparison of enzymes from strains of wine yeast and their related yeasts on electrophoresis. J. Inst. Enol. Vitic. Yamanashi Univ. 17, 11–19 (1982)Google Scholar
  38. 38.
    Yarrow, D. &T. Nakase: DNA base compositions of the genus Saccharomyces. Antonie van Leeuwenhoek 41, 81–88 (1975)PubMedCrossRefGoogle Scholar

Copyright information

© Carlsberg Laboratory 1983

Authors and Affiliations

  • Mogens Bohl Pedersen
    • 1
    • 2
  1. 1.Department of PhysiologyCarlsberg LaboratoryCopenhagen, Valby
  2. 2.Department of Brewing ChemistryCarlsberg Research LaboratoryCopenhagen, Valby

Personalised recommendations