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. 1988 Mar;8(3):1027–1035. doi: 10.1128/mcb.8.3.1027

Constitutive expression of the maltose fermentative enzymes in Saccharomyces carlsbergensis is dependent upon the mutational activation of a nonessential homolog of MAL63.

R A Dubin 1, M J Charron 1, S R Haut 1, R B Needleman 1, C A Michels 1
PMCID: PMC363245  PMID: 2835655

Abstract

Maltose fermentation in Saccharomyces carlsbergensis is dependent upon the MAL6 locus. This complex locus is composed of the MAL61 and MAL62 genes, which encode maltose permease and maltase, respectively, and a third gene, MAL63, which codes for a trans-acting positive regulatory product. In wild-type strains, expression of the MAL61 and MAL62 mRNAs and proteins is induced by maltose and induction is dependent upon the MAL63 gene. Mutants constitutively expressing the MAL61 and MAL62 gene products have been isolated in mal63 backgrounds, and the mutations which have been analyzed map to a fourth MAL6-linked gene, MAL64. Cloning and characterization of this new gene are described in this report. The results revealed that the MAL64-C alleles present in constitutive strains encode a trans-acting positive function required for constitutive expression of the MAL61 and MAL62 gene products. In inducible strains, the MAL64 gene is dispensable, as deletion of the gene had no effect on maltose fermentation or maltose-regulated induction. MAL64 encoded transcripts of 2.0 and 1.4 kilobase pairs. While both MAL64 mRNAs were constitutively expressed in constitutive strains, they were maltose inducible in wild-type strains and induction was dependent upon the MAL63 gene. The MAL63 and MAL64 genes are at least partially structurally homologous, suggesting that they control MAL61 and MAL62 transcript accumulation by similar mechanisms.

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Selected References

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  1. Barnett J. A. The utilization of sugars by yeasts. Adv Carbohydr Chem Biochem. 1976;32:125–234. doi: 10.1016/s0065-2318(08)60337-6. [DOI] [PubMed] [Google Scholar]
  2. Boeke J. D., LaCroute F., Fink G. R. A positive selection for mutants lacking orotidine-5'-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet. 1984;197(2):345–346. doi: 10.1007/BF00330984. [DOI] [PubMed] [Google Scholar]
  3. Broach J. R., Strathern J. N., Hicks J. B. Transformation in yeast: development of a hybrid cloning vector and isolation of the CAN1 gene. Gene. 1979 Dec;8(1):121–133. doi: 10.1016/0378-1119(79)90012-x. [DOI] [PubMed] [Google Scholar]
  4. Cameron J. R., Loh E. Y., Davis R. W. Evidence for transposition of dispersed repetitive DNA families in yeast. Cell. 1979 Apr;16(4):739–751. doi: 10.1016/0092-8674(79)90090-4. [DOI] [PubMed] [Google Scholar]
  5. Charron M. J., Dubin R. A., Michels C. A. Structural and functional analysis of the MAL1 locus of Saccharomyces cerevisiae. Mol Cell Biol. 1986 Nov;6(11):3891–3899. doi: 10.1128/mcb.6.11.3891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Charron M. J., Michels C. A. The constitutive, glucose-repression-insensitive mutation of the yeast MAL4 locus is an alteration of the MAL43 gene. Genetics. 1987 May;116(1):23–31. doi: 10.1093/genetics/116.1.23. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cohen J. D., Goldenthal M. J., Buchferer B., Marmur J. Mutational analysis of the MAL1 locus of Saccharomyces: identification and functional characterization of three genes. Mol Gen Genet. 1984;196(2):208–216. doi: 10.1007/BF00328052. [DOI] [PubMed] [Google Scholar]
  8. Cohen J. D., Goldenthal M. J., Chow T., Buchferer B., Marmur J. Organization of the MAL loci of Saccharomyces. Physical identification and functional characterization of three genes at the MAL6 locus. Mol Gen Genet. 1985;200(1):1–8. doi: 10.1007/BF00383304. [DOI] [PubMed] [Google Scholar]
  9. Dubin R. A., Needleman R. B., Gossett D., Michels C. A. Identification of the structural gene encoding maltase within the MAL6 locus of Saccharomyces carlsbergensis. J Bacteriol. 1985 Nov;164(2):605–610. doi: 10.1128/jb.164.2.605-610.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dubin R. A., Perkins E. L., Needleman R. B., Michels C. A. Identification of a second trans-acting gene controlling maltose fermentation in Saccharomyces carlsbergensis. Mol Cell Biol. 1986 Aug;6(8):2757–2765. doi: 10.1128/mcb.6.8.2757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Federoff H. J., Cohen J. D., Eccleshall T. R., Needleman R. B., Buchferer B. A., Giacalone J., Marmur J. Isolation of a maltase structural gene from Saccharomyces carlsbergensis. J Bacteriol. 1982 Mar;149(3):1064–1070. doi: 10.1128/jb.149.3.1064-1070.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hall B. G., Yokoyama S., Calhoun D. H. Role of cryptic genes in microbial evolution. Mol Biol Evol. 1983 Dec;1(1):109–124. doi: 10.1093/oxfordjournals.molbev.a040300. [DOI] [PubMed] [Google Scholar]
  13. Hong S. H., Marmur J. Primary structure of the maltase gene of the MAL6 locus of Saccharomyces carlsbergensis. Gene. 1986;41(1):75–84. doi: 10.1016/0378-1119(86)90269-6. [DOI] [PubMed] [Google Scholar]
  14. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Khan N. A., Eaton N. R. Genetic control of maltase formation in yeast. I. Strains producing high and low basal levels of enzyme. Mol Gen Genet. 1971;112(4):317–322. doi: 10.1007/BF00334433. [DOI] [PubMed] [Google Scholar]
  16. Needleman R. B., Kaback D. B., Dubin R. A., Perkins E. L., Rosenberg N. G., Sutherland K. A., Forrest D. B., Michels C. A. MAL6 of Saccharomyces: a complex genetic locus containing three genes required for maltose fermentation. Proc Natl Acad Sci U S A. 1984 May;81(9):2811–2815. doi: 10.1073/pnas.81.9.2811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Needleman R., Eaton N. R. Selection of yeast mutants constitutive for maltase synthesis. Mol Gen Genet. 1974;133(2):135–140. doi: 10.1007/BF00264834. [DOI] [PubMed] [Google Scholar]
  18. Ng R., Abelson J. Isolation and sequence of the gene for actin in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3912–3916. doi: 10.1073/pnas.77.7.3912. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Orr-Weaver T. L., Szostak J. W., Rothstein R. J. Genetic applications of yeast transformation with linear and gapped plasmids. Methods Enzymol. 1983;101:228–245. doi: 10.1016/0076-6879(83)01017-4. [DOI] [PubMed] [Google Scholar]
  20. Panek A. D., Sampaio A. L., Braz G. C., Baker S. J., Mattoon J. R. Genetic and metabolic control of trehalose and glycogen synthesis. New relationships between energy reserves, catabolite repression and maltose utilization. Cell Mol Biol Incl Cyto Enzymol. 1979;25(5):345–354. [PubMed] [Google Scholar]
  21. Rothstein R. J. One-step gene disruption in yeast. Methods Enzymol. 1983;101:202–211. doi: 10.1016/0076-6879(83)01015-0. [DOI] [PubMed] [Google Scholar]
  22. Van Wijk R., Ouwehand J., van den Bos T., Koningsberger V. V. Induction and catabolite repression of alpha-glucosidase synthesis in protoplasts of Saccharomyces carlsbergensis. Biochim Biophys Acta. 1969 Jul 22;186(1):178–191. doi: 10.1016/0005-2787(69)90501-2. [DOI] [PubMed] [Google Scholar]
  23. Zimmermann F. K., Eaton N. R. Genetics of induction and catabolite repression of Maltese synthesis in Saccharomyces cerevisiae. Mol Gen Genet. 1974;134(3):261–272. doi: 10.1007/BF00267720. [DOI] [PubMed] [Google Scholar]
  24. de Kroon R. A., Koningsberger V. V. An inducible transport system for alpha-glucosides in protoplasts of Saccharomyces carlsbergensis. Biochim Biophys Acta. 1970 Apr 15;204(2):590–609. doi: 10.1016/0005-2787(70)90178-4. [DOI] [PubMed] [Google Scholar]
  25. ten Berge A. M., Zoutewelle G., Needleman R. B. Regulation of maltose fermentation in Saccharomyces carlsbergensis. 3. Constitutive mutations at the MAL6-locus and suppressors changing a constitutive phenotype into a maltose negative phenotype. Mol Gen Genet. 1974;131(2):113–121. doi: 10.1007/BF00266147. [DOI] [PubMed] [Google Scholar]
  26. ten Berge A. M., Zoutewelle G., van de Poll K. W., Bloemers H. P. Regulation of maltose fermentation in Saccharomyces carlsbergensis. II. Properties of a constitutive MAL6-mutant. Mol Gen Genet. 1973 Sep 5;125(2):139–146. doi: 10.1007/BF00268867. [DOI] [PubMed] [Google Scholar]
  27. ten Berge A. M., Zoutewelle G., van de Poll K. W. Regulation of maltose fermentation in Saccharomyces carlsbergensis. I. The function of the gene MAL6, as recognized by mal6-mutants. Mol Gen Genet. 1973 Jul 2;123(3):233–246. doi: 10.1007/BF00271242. [DOI] [PubMed] [Google Scholar]

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