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. 1997 Aug;146(4):1287–1298. doi: 10.1093/genetics/146.4.1287

Constitutive Mutations of the Saccharomyces Cerevisiae Mal-Activator Genes Mal23, Mal43, Mal63, and Mal64

A W Gibson 1, L A Wojciechowicz 1, S E Danzi 1, B Zhang 1, J H Kim 1, Z Hu 1, C A Michels 1
PMCID: PMC1208075  PMID: 9258674

Abstract

We report the sequence of several MAL-activator genes, including inducible, constitutive, and noninducible alleles of MAL23, MAL43, MAL63, and mal64. Constitutive alleles of MAL23 and MAL43 vary considerably from inducible alleles in their C-terminal domain, with many of the alterations clustered and common to both alleles. The 27 alterations from residues 238-461 of Mal43-C protein are sufficient for constitutivity, but the minimal number of alterations needed for the constitutive phenotype could not be determined. The sequence of mal64, a nonfunctional homologue of MAL63, revealed that Mal64p is 85% identical to Mal63p. Two mutations that activate mal64 and cause constitutivity are nonsense mutations resulting in truncated proteins of 306 and 282 residues. We conclude that the C-terminal region of the MAL-activator, from residues 283-470, contains a maltose-responsive negative regulatory domain, and that extensive mutation or deletion of the entire region causes loss of the negative regulatory function. Additionally, certain sequence elements in the region appear to be necessary for efficient induction of the full-length Mal63 activator protein. These studies highlight the role of ectopic recombination as an important mechanism of mutagenesis of the telomere-associated family of MAL loci.

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Selected References

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  1. Chang Y. S., Dubin R. A., Perkins E., Forrest D., Michels C. A., Needleman R. B. MAL63 codes for a positive regulator of maltose fermentation in Saccharomyces cerevisiae. Curr Genet. 1988 Sep;14(3):201–209. doi: 10.1007/BF00376740. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. 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]
  4. Charron M. J., Michels C. A. The naturally occurring alleles of MAL1 in Saccharomyces species evolved by various mutagenic processes including chromosomal rearrangement. Genetics. 1988 Sep;120(1):83–93. doi: 10.1093/genetics/120.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cheng Q., Michels C. A. The maltose permease encoded by the MAL61 gene of Saccharomyces cerevisiae exhibits both sequence and structural homology to other sugar transporters. Genetics. 1989 Nov;123(3):477–484. doi: 10.1093/genetics/123.3.477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dubin R. A., Charron M. J., Haut S. R., Needleman R. B., Michels C. A. Constitutive expression of the maltose fermentative enzymes in Saccharomyces carlsbergensis is dependent upon the mutational activation of a nonessential homolog of MAL63. Mol Cell Biol. 1988 Mar;8(3):1027–1035. doi: 10.1128/mcb.8.3.1027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Elledge S. J., Davis R. W. A family of versatile centromeric vectors designed for use in the sectoring-shuffle mutagenesis assay in Saccharomyces cerevisiae. Gene. 1988 Oct 30;70(2):303–312. doi: 10.1016/0378-1119(88)90202-8. [DOI] [PubMed] [Google Scholar]
  10. Feuermann M., Charbonnel L., De Montigny J., Bloch J. C., Potier S., Souciet J. L. Sequence of a 9.8 kb segment of yeast chromosome II including the three genes of the MAL3 locus and three unidentified open reading frames. Yeast. 1995 Jun 15;11(7):667–672. doi: 10.1002/yea.320110707. [DOI] [PubMed] [Google Scholar]
  11. Han E. K., Cotty F., Sottas C., Jiang H., Michels C. A. Characterization of AGT1 encoding a general alpha-glucoside transporter from Saccharomyces. Mol Microbiol. 1995 Sep;17(6):1093–1107. doi: 10.1111/j.1365-2958.1995.mmi_17061093.x. [DOI] [PubMed] [Google Scholar]
  12. Hu Z., Nehlin J. O., Ronne H., Michels C. A. MIG1-dependent and MIG1-independent glucose regulation of MAL gene expression in Saccharomyces cerevisiae. Curr Genet. 1995 Aug;28(3):258–266. doi: 10.1007/BF00309785. [DOI] [PubMed] [Google Scholar]
  13. Johnston S. A., Hopper J. E. Isolation of the yeast regulatory gene GAL4 and analysis of its dosage effects on the galactose/melibiose regulon. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6971–6975. doi: 10.1073/pnas.79.22.6971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Kim J., Michels C. A. The MAL63 gene of Saccharomyces encodes a cysteine-zinc finger protein. Curr Genet. 1988 Oct;14(4):319–323. doi: 10.1007/BF00419988. [DOI] [PubMed] [Google Scholar]
  16. Levine J., Tanouye L., Michels C. A. The UAS(MAL) is a bidirectional promotor element required for the expression of both the MAL61 and MAL62 genes of the Saccharomyces MAL6 locus. Curr Genet. 1992 Sep;22(3):181–189. doi: 10.1007/BF00351724. [DOI] [PubMed] [Google Scholar]
  17. Michels C. A., Read E., Nat K., Charron M. J. The telomere-associated MAL3 locus of Saccharomyces is a tandem array of repeated genes. Yeast. 1992 Aug;8(8):655–665. doi: 10.1002/yea.320080809. [DOI] [PubMed] [Google Scholar]
  18. Mézard C., Pompon D., Nicolas A. Recombination between similar but not identical DNA sequences during yeast transformation occurs within short stretches of identity. Cell. 1992 Aug 21;70(4):659–670. doi: 10.1016/0092-8674(92)90434-e. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Needleman R. Control of maltase synthesis in yeast. Mol Microbiol. 1991 Sep;5(9):2079–2084. doi: 10.1111/j.1365-2958.1991.tb02136.x. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Papa F. R., Hochstrasser M. The yeast DOA4 gene encodes a deubiquitinating enzyme related to a product of the human tre-2 oncogene. Nature. 1993 Nov 25;366(6453):313–319. doi: 10.1038/366313a0. [DOI] [PubMed] [Google Scholar]
  23. Rogers S., Wells R., Rechsteiner M. Amino acid sequences common to rapidly degraded proteins: the PEST hypothesis. Science. 1986 Oct 17;234(4774):364–368. doi: 10.1126/science.2876518. [DOI] [PubMed] [Google Scholar]
  24. Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. WINGE O., ROBERTS C. The polymeric genes for maltose fermentation in yeasts, and their mutability. Cr Trav Lab Carlsberg Ser Physiol. 1950;25(2):35–89. [PubMed] [Google Scholar]
  26. Wang J., Needleman R. Removal of Mig1p binding site converts a MAL63 constitutive mutant derived by interchromosomal gene conversion to glucose insensitivity. Genetics. 1996 Jan;142(1):51–63. doi: 10.1093/genetics/142.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. 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]
  29. 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]
  30. 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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