Abstract
Many sets of genes in Saccharomyces cerevisiae are divergently transcribed, but at present there are no vectors generally available for the simultaneous analysis of divergent transcription from these promoters. In the present study MEL1 and lacZ were used to construct a vector capable of measuring the divergent expression initiated by the MAL6T-MAL6S bi-directional promoter. Our observations demonstrate that the expression of both reporter genes was regulated in a similar fashion to the native MAL6T and MAL6S genes, and that induction was dependent upon the presence of a functional MALR activator gene. The results confirmed that the MAL6T-MAL6S promoter was co-ordinately regulated, repressed by glucose, induced by maltose, and that basal expression was more active in the MAL6S direction than in the MAL6T direction.
Similar content being viewed by others
Explore related subjects
Discover the latest articles and news from researchers in related subjects, suggested using machine learning.References
Astell CR, Ahlstrom-Jonasson L, Smith M, Tatchell K, Nasmyth KA, Hall BD (1981) The sequence of the DNAs encoding for the mating-type loci of Saccharomyces cerevisiae. Cell 27:15–23
Barnett JA (1976) The utilisation of sugars by yeast. Adv Carbohydr Chem Biochem 32:125–134
Beck CF, Warren RAJ (1988) Divergent promoters, a common form of gene organisation. Microbiol Rev 318–326
Bradford M (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilising the principle of protein-dye binding. Anal Biochem 72:248–254
Briza P, Breitenbach M, Ellinger A, Segall J (1990) Isolation of two developmentally regulated genes involved in spore-wall maturation in Saccharomyces cerevisiae. Genes Dev 4:1775–1789
Chang YS, Dubin RA, Perkins E, Forrest D, Michels CA (1988) MAL63 codes for a positive regulator of maltose fermentation in Saccharomyces cerevisiae. Curr Genet 14:201–209
Cohen JD, Goldenthal JD, Chow T, Buchferer B, Marmur J (1985) Mutational analysis of the MAL1 locus of Saccharomyces: identification and functional characterisation of three genes. Mol Gen Genet 200:1–8
Dubin RA, Perkins EL, Needleman RB, Michels CA (1986) Identification of a second trans-acting gene controlling maltose fermentation in Saccharomyces carlsbergensis. Mol Cell Biol 6:2757–2765
Dubin RA, Charron MJ, Haut SR, Needleman RB, Michels CA (1988) Constitutive expression of the maltose fermentative enzymes in Saccharomyces carlsbergensis is dependent upon the mutationl activation of a non-essential homology of MAL63. Mol Cell Biol 8:1027–1035
Dujon B, Alexandraki D, Andre B, et al. (1994) Complete DNA sequence of yeast chromosome XI. Nature 369:371–378
Goldenthal MJ, Vanoni M, Buchferer B, Marmur J (1987) Regulation of MAL gene expression in yeast: gene dosage effects. Mol Gen Genet 209:508–517
Hong SH, Marmur J (1987) Primary structure of the maltase gene of the MAL6 locus of Saccharomyces carlsbergensis. Gene 41:75–84
Hong SH, Marmur J (1987) Upstream regulatory regions controlling the expression of the yeast maltase gene. Mol Cell Biol 7:2477–2483
Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:33–41
Johnston M, Davis R (1984) Sequences that regulate the divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae. Mol Cell Biol 4:1440–1448
Kew OM, Douglas HC (1976) Genetic co-regulation of galactose and melibiose utilization in Saccharomyces. J Bacteriol 125:33–41
Levine J, Tanouye L, Michels CA (1992) The UASMAL is a bidirectional promoter element required for the expression of both the MAL61 and MAL62 genes of the Saccharomyces MAL6 locus. Curr Genet 22:181–189
Liljestrom PL (1985) The nucleotide sequence of the yeast MEL1 gene. Nucleic Acids Res 13:7257–7268
Lowry CV, Weiss JL, Walthall DA, Zitomer RS (1983) Modulator sequences mediate the oxygen regulation of CYC1 and a neighboring gene in yeast. Proc Natl Acad Sci USA 80:151–155
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Myers AM, Tzagaloff A, Kinney DM, Lusty CJ (1986) Yeast shuttle and integrative vectors with multiple cloning sites suitable for the construction of lacZ fusions. Gene 45:299–310
Needleman R (1991) Control of maltase synthesis in yeast. Mol Microbiol 5:2079–2084
Needleman RB, Kaback DB, Dubin RA, Perkins EL, Rosenberg NG, Sutherland KA, Forrest DB, Michels CA (1984) MAL6 of Saccharomyces: a complex locus containing three genes required for maltose fermentation. Proc Natl Acad Sci USA 81:2811–2815
Oliver SG, van der Art QJM, Agostoni-Carbone ML, et al. (1992) The complete sequence of yeast chromosome III. Nature 357:38–46
Osley MA, Gould J, Kim S, Kane M, Hereford L (1986) Identification of sequences in a yeast hitone promoter involved in periodic transcription. Cell 45:537–544
Post-Beittenmiller MA, Hamilton RW, Hopper JE (1984) Regulation of basal and induced levels of the MEL1 transcript in Saccharomyces cerevisiae. Mol Cell Biol 4:1238–1245
Rodicio R, Zimmermann (1985) Cloning of maltase regulatory genes in Saccharomyces cerevisiae. Curr Genet 9:539–545
Ronald SL, Kropinski AM, Farinha MA (1990) Construction of broad-host-range vectors for the selection of divergent promoters. Gene 90:145–148
Schmitt EM, Brown TA, Trumpower L (1990) A rapid and simple method for preparation of RNA from Saccharomyces cerevisiae. Nucleic Acids Research 18:3091–3092
Schneider K, Beck CF (1986) Promoter vectors for the analysis of divergently arranged promoters. Gene 42:37–48
Struhl K, Davis RW (1981) Promoter mutants of the yeast his3 gene. J Mol Biol 152:535–552
Struhl K, Stinchcomb DT, Scherer S, Davis RW (1979) High frequency transformation of yeast: autonomous replication of hybrid DNA molecules. Proc Natl Acad Sci USA 76:1035–1039
Sumner-Smith M, Bozzato RP, Skipper N, Davies RW, Hopper JE (1985) Analysis of the inducible MEL1 gene of Saccharomyces carlsbergensis and its secreted product, alpha-galactosidase (melibiase). Gene 36:333–340
West RW Jr, Yocum RR, Ptashne M (1984) The GAL1-GAL10 divergent promoter region of Saccharomyces cerevisiae contains negative control elements in addition to functionally separate and possibly overlapping upstream activating sequences. Mol Cell Biol 4:2467–2478
Yao B, Sollitti P, Zhang XL, Marmur J (1994) Shared control of maltose induction and catabolite repression of the MAL structural genes in Saccharomyces. Mol Gen Genet 243:622–630
Author information
Authors and Affiliations
Additional information
Communicated by M. S. Esposito
Rights and permissions
About this article
Cite this article
Bell, P.J.L., Bissinger, P.H., Evans, R.J. et al. A two-reporter gene system for the analysis of bi-directional transcription from the divergent MAL6T-MAL6S promoter in Saccharomyces cerevisiae . Curr Genet 28, 441–446 (1995). https://doi.org/10.1007/BF00310813
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00310813


