MAL6 of Saccharomyces: a complex genetic locus containing three genes required for maltose fermentation.

Needleman, Richard; Kaback, David B.; Dubin, Robert; Perkins, Edward L.; Rosenberg, Nurit; Sutherland, Karen; Forrest, Doug; Michels, Corinne A.

doi:10.1073/pnas.81.9.2811

Cited by 114 publications

(90 citation statements)

References 23 publications

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“…3) contains a fragment internal to the MAL62 gene. This probe detected a 1.9-kb maltose-inducible transcript in MAL6 strains (11,22). Plasmid pD-5 also detected a 1.9-kb maltose-inducible transcript in the MALl strain 600-1B (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Plasmid DNA and yeast DNA and RNA isolations as well as Southern and Northern analyses were carried out as previously described except that polyadenylated [poly(A)+] RNA was isolated following a single pass through oligo(dT)-cellulose (11,22).…”

Section: Methodsmentioning

confidence: 99%

“…Federoff et al (13) cloned a segment of DNA from Saccharomyces carlsbergensis CB11 that was shown to contain the MAL6 locus (23). This cloned fragment contains three transcribed regions which we refer to as MAL61, MAL62, and MAL63 (22). All three genes are required for maltose fermentation.…”

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confidence: 99%

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Structural and functional analysis of the MAL1 locus of Saccharomyces cerevisiae.

Charron¹,

Dubin²,

Michels³

1986

Mol. Cell. Biol.

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We describe the isolation of a 22.6-kilobase fragment of DNA containing the MALI locus of Saccharomyces cerevisiae. Our results demonstrate that the MALI locus, like the MAL6 locus, is a complex locus containing three genes. These genes were organized similarly to their MAL6 counterparts. We refer to them as MALI], MAL12, and MAL13 and show that they are functionally homologous to the MAL61 (encoding maltose permease), MAL62 (encoding maltase), and MAL63 (encoding the positive regulator) genes of the MAL6 locus. Transcription from each of the three genes was analyzed in a strain carrying the undisrupted MALI locus and in strains carrying single disruptions in each of the MALI genes. The MALI and MAL6 loci were found to be highly sequence homologous and conserved throughout the region containing these three genes. The strain used to isolate the MALI locus also carried the tightly linked SUCI gene. The SUCI gene was found to be located on the same 22.6-kilobase fragment containing the MALI locus and 5 kilobases from the 3' end of the MAL12 gene. The meaning of these results with regard to the mechanism of regulation of maltose fermentation is discussed.The fermentation of maltose by Saccharomyces spp. requires the presence of at least one of a series of five unlinked loci: MALI, MAL2, MAL3, MAI4, or MAL6 (1). Several standard maltose-fermenting laboratory strains have been genetically analyzed and shown to carry one dominant MAL locus and one or two additional, partially functional, MAL loci. Physical analysis of these strains revealed extensive sequence homology between the cloned MAL6 locus and the other MAL loci (7,17,18,23).The maltose-fermentative enzymes maltase and maltose permease are subject to both maltose induction and glucose repression. In the presence of maltose, both of these enzymes are induced approximately 30-fold, while in the noninduced state, only low basal levels of these enzymes are present (10,26). It has been postulated that a positive regulatory function is required for induction and that a gene encoding this regulatory function maps to each of the dominant MAL loci (28,29). More recent genetic analysis of maltose fermentation has revealed other functions encoded by the MAL loci which are required for fermentation (8, 11, lla, 14, 22; Chang et al., submitted for publication). Federoff et al. (13) cloned a segment of DNA from Saccharomyces carlsbergensis CB11 that was shown to contain the MAL6 locus (23). This cloned fragment contains three transcribed regions which we refer to as MAL61, MAL62, and MAL63 (22). All three genes are required for maltose fermentation. Gene disruption experiments at the MAL6 locus have confirmed these results (11; Chang et al., submitted). MAL63 has been shown to encode the positive regulatory gene product required for induction of maltase and maltose permease (Chang et al., submitted). Gene disruption experiments have shown that MAL62 is the structural gene encoding maltase (11). MAL61 is believed to code for maltose permease (9; Chang et al., submitted). Mut...

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Structural and functional analysis of the MAL1 locus of Saccharomyces cerevisiae.

Charron¹,

Dubin²,

Michels³

1986

Mol. Cell. Biol.

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“…Strain NCYC 1324 also possessed sequences corresponding to the MAL2 locus. The MAL6 locus, which has been the subject of extensive investigation 8,25,35,43,53,63 , was not detected by the MAL31 probe in the strains studied. In addition to the reactive sequences that could be assigned to specific loci, hybridisation to other bands was observed.…”

Section: (Ixigxmsr Sj Kiriw Mrzspzih Mr Qepxswi Erh Wygvswi Yxmpmwexmsrmentioning

confidence: 99%

Expression Patterns of Genes and Enzymes Involved in Sugar Catabolism in IndustrialSaccharomyces cerevisiaeStrains Displaying Novel Fermentation Characteristics

Meneses

Jiranek

2002

Journal of the Institute of Brewing

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The utilisation of maltose or sucrose by a selection of nine brewing, baking or laboratory strains of Saccharomyces cerevisiae was either repressible, constitutive or absent. Overall fermentation rate showed a good correlation with maximum specific maltose transport rate (R 2 = 0.79), but a poor correlation with maximum maltase activity (R 2 = 0.34), implying that transport rather than hydrolysis of maltose was a rate-limiting step determining fermentation performance. The genetic basis for differences seen between the strains in terms of fermentation kinetics was investigated. All strains were found to possess genomic sequences detectable with probes for the MAL11 (AGT1), MAL31 permease genes and the SUC2 invertase gene, however, the loci present and the pattern of expression of these and other MAL genes varied widely. From comparisons of sugar utilisation and gene expression patterns, constitutive maltose utilisation in strain NCYC 1681 was best explained by AGT1 expression. Such expression of AGT1 was not, however, mirrored by changes in the pattern of expression of the transcriptional activator(s) (MALx3). Patterns of sucrose utilisation were poorly predicted by the kinetics of SUC2 gene expression, indicating that other SUC loci may be of greater importance.

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“…Total RNA from S. cerevisiae was isolated by the method of Needleman et al (31). RNA was separated in 1.0% agarose gels and transferred to nylon membranes as described previously (7).…”

Section: Methodsmentioning

confidence: 99%

Proline biosynthesis in Saccharomyces cerevisiae: analysis of the PRO3 gene, which encodes delta 1-pyrroline-5-carboxylate reductase

Brandriss

Falvey

1992

J Bacteriol

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The PRO3 gene of Saccharomyces cerevisiae encodes the 286-amino-acid protein AW-pyrroline-5-carboxylate reductase [L-proline:NAD(P+) 5-oxidoreductase; EC 1.5.1.21, which catalyzes the final step in proline biosynthesis. The protein has substantial similarity to the pyrroline carboxylate reductases of diverse bacterial species, soybean, and humans. Using RNA hybridization and measurements of enzyme activity, we have determined that the expression of the PRO3 gene appears to be constitutive. It is not repressed by the pathway end product (proline), induced by the initial substrate (glutamate), or regulated by the general control system. Its expression is not detectably altered when cells are grown in a wide range of nitrogen sources or when glycerol and ethanol replace glucose as the carbon source. The possibility that this enzyme has other functions in addition to proline biosynthesis is discussed.In the yeast Saccharomyces cerevisiae, the pathway of proline biosynthesis was established by characterization of proline-requiring mutants and interspecies complementation of Escherichia coli and Salmonella typhimurium proline auxotrophs by the cloned yeast genes (6,38 In addition to the formation of proline required for protein synthesis, the final step in the proline biosynthetic pathway generates NADP+, which is believed to stimulate the production of ribose-5-phosphate via the pentose phosphate shunt in human erythrocytes (42) and in soybean nodules (22). This stimulation results in an increase in 5-phosphoribosyl 1-pyrophosphate, leading to an increase in de novo purine biosynthesis. Because of the variety of metabolic interconversions of proline and P5C, these compounds link the tricarboxylic acid and urea cycles with amino acid and purine biosynthesis in a diversity of cell types.With this report, we begin a molecular analysis of the S. cerevisiae gene encoding P5C reductase, the enzyme that catalyzes the final step of proline biosynthesis. Because arginine catabolism and proline biosynthesis converge at P5C (10), P5C reductase is both a proline-biosynthetic and an arginine-degradative enzyme. A biochemical analysis of the purified protein (26, 27) and its cytosolic localization (11) * Corresponding author.were reported a decade ago. pro3 mutants require proline for growth, lack P5C reductase activity, and have the peculiar property of being unable to grow on rich, or completely supplemented, minimal media (6,38). Until now, very little has been published concerning the regulation of this gene or its product. Here, we report the DNA sequence of the PRO3 gene and show that its deduced amino acid sequence has substantial similarity to that of P5C reductases in diverse bacterial species, soybean, and humans. We also measured PRO3 gene expression and P5C reductase activity as a function of growth on various nitrogen and carbon sources, the presence of proline or glutamate in the medium, and its sensitivity to proline starvation and the general control system that regulates the expression of many amino acidbiosynthet...

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MAL6 of Saccharomyces: a complex genetic locus containing three genes required for maltose fermentation.

Cited by 114 publications

References 23 publications

Structural and functional analysis of the MAL1 locus of Saccharomyces cerevisiae.

Structural and functional analysis of the MAL1 locus of Saccharomyces cerevisiae.

Expression Patterns of Genes and Enzymes Involved in Sugar Catabolism in IndustrialSaccharomyces cerevisiaeStrains Displaying Novel Fermentation Characteristics

Proline biosynthesis in Saccharomyces cerevisiae: analysis of the PRO3 gene, which encodes delta 1-pyrroline-5-carboxylate reductase

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