Molecular and General Genetics MGG

, Volume 191, Issue 3, pp 366–371 | Cite as

Identification and physical characterization of yeast maltase structural genes

  • T. Chow
  • M. J. Goldenthal
  • J. D. Cohen
  • M. Hegde
  • J. Marmur
Article

Summary

Each of at least five unlinked MAL loci (MAL1 through MAL4 and MAL6) on the yeast genome controls the ability to synthesize an inducible α-D-glucosidase (maltase). A subcloned fragment of the coding sequence of the MAL6 maltase structural gene was used as a hybridization probe to investigate the physical structure of the family of MAL structural genes in the genomes of different Saccharomyces strains. Mal+ strains, each carrying a genetically defined MAL locus, were crossed with a Mal- strain and the segregation behavior of the functional locus and of sequences complementary to the maltase structural gene at that locus analyzed. The maltase structural gene sequences of each MAL locus were detected by Southern blot hybridization using BamH1 digests of genomic DNA of the meiotic products. This restriction enzyme was previously shown to cleave outside the confines of the MAL6 locus.

The results of such experiments indicate that each MAL locus encompasses at least one maltase structural gene sequence homologous to that of MAL6, that yeast strains that lack functional MAL loci may or may not contain the corresponding maltase structural gene sequence, that the MAL1 maltase structural gene sequence or one of its alleles can be detected in all laboratory yeast strains examined and that each MAL locus can be identified as a characteristic BamH1 fragment of genomic DNA which includes a maltase structural gene.

Yeast strains vary in the number of maltase structural gene sequences that they carry. By using the approach described in this report, the ones corresponding to the different functional MAL loci and residing within a BamH1 generated restriction fragment can be identified.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Barnett JA (1976) The utilization of sugars by yeast. Adv Carbohydr Chem Biochem 32:125–234Google Scholar
  2. Carlson M, Osmond BC, Botstein D (1981) SUC genes of yeast: a dispersed gene family. Cold Spring Harbor Symp Quant Biol 45(B):799–803Google Scholar
  3. Cryer DR, Eccleshall TR, Marmur J (1975) Isolation of yeast DNA. Methods Cell Biol 12:39–44Google Scholar
  4. Federoff HJ, Cohen JD, Eccleshall TR, Needleman RB, Buchferer BA, Giacolone J, Marmur J (1982) Isolation of a maltase structural gene from Saccharomyces carlsbergensis. J Bacteriol 116:1064–1066Google Scholar
  5. Goldenthal MJ, Cohen JD, Marmur J (1983) Isolation and characterization of a maltose transport mutant in Saccharomyces cerevisiae. Curr Genet 7:195–199Google Scholar
  6. Holland JP, Holland MJ (1980) Structural comparison of two nontandemly repeated yeast glyceraldehyde 3-phosphate dehydrogenase genes. J Biol Chem 255:2596–2605Google Scholar
  7. Messing J (1981) M13mp2 and derivatives; a molecular cloning system for DNA sequencing, strand specific hybridization and in vitro mutagenesis. In: Walton A (ed) Third Cleveland Symposium on Macromolecules: Recombinant DNA. Elsevier, Amsterdam, p 143–153Google Scholar
  8. Mowshowitz DB (1981) The effects of three different MAL loci on the regulation of maltase synthesis in yeast. Genetics 98:713–728Google Scholar
  9. Naumov GI (1981) Comparative genetics in yeast. V. Complementation in the MAL locus in Saccharomyces which do not utilize maltose. Gentika 7:141–148Google Scholar
  10. Naumov GI (1976) Comparative genetics of yeast. XVI. Genes for maltose fermentation in Saccharomyces carlsbergensis. Genetika 12:87–100Google Scholar
  11. Rogers D, Lemire JM, Bostian KA (1982) Acid phosphatase polypeptides in Saccharomyces cerevisiae are encoded by a differentially regulated multigene family. Proc Natl Acad Sci USA 79:2157–2161Google Scholar
  12. Sherman F, Fink GR, Lawrence CW (1972) Laboratory manual for methods in yeast genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor NYGoogle Scholar
  13. Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517Google Scholar
  14. ten Berge AMA (1972) Genes for the fermentation of maltose and α-methyl glucoside in Saccharomyces carlsbergensis. Mol Gen Genet 115:80–88Google Scholar
  15. ten Berge AMA, Zoutevelle G, Van de Poll W (1975) Regulation of maltose fermentation in Saccharomyces carlsbergensis. I. The function of the gene MAL6 as recognized by mal6 mutants. Mol Gen Genet 123:233–246Google Scholar
  16. Wahl GM, Stern M, Stark G (1979) Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl paper and rapid hybridization using dextran sulfate. Proc Natl Acad Sci USA 76:3683–3687Google Scholar

Copyright information

© Springer-Verlag 1983

Authors and Affiliations

  • T. Chow
    • 1
  • M. J. Goldenthal
    • 1
  • J. D. Cohen
    • 1
  • M. Hegde
    • 1
  • J. Marmur
    • 1
  1. 1.Department of BiochemistryAlbert Einstein College of MedicineBronxUSA

Personalised recommendations