Repeated Family of Genes Controlling Maltose Fermentation in Saccharomyces carlsbergensis

1983 ◽  
Vol 3 (5) ◽  
pp. 796-802
Author(s):  
Richard B. Needleman ◽  
Corinne Michels
Keyword(s):  
Genetic Analysis ◽  
Structural Gene ◽  
Regulatory Gene ◽  
Regulatory Protein ◽  
Physical Analysis ◽  
Saccharomyces Carlsbergensis ◽  
Maltose Permease ◽  
Maltose Fermentation ◽  
Complex Locus ◽  
Dna Fragment

Maltose fermentation in Saccharomyces spp. requires the presence of any one of five unlinked genes: MAL1, MAL2, MAL3, MAL4 , or MAL6. Although the genes are functionally equivalent, their natures and relationships to each other are not known. At least three proteins are necessary for maltose fermentation: maltase, maltose permease, and a regulatory protein. The MAL genes may code for one or more of these proteins. Recently a DNA fragment containing a maltase structural gene has been cloned from a MAL6 strain, CB11, to produce plasmid pMAL9-26. We have conducted genetic and physical analyses of strain CB11. The genetic analysis has demonstrated the presence of two cryptic MAL genes in CB11, MAL1g and MAL3g (linked to MAL1 and to MAL3 , respectively), in addition to the MAL6 locus. The physical analysis, which used a subclone of plasmid pMAL9-26 as a probe, detected three Hin dIII genomic fragments with homology to the probe. Each fragment was shown to be linked to one of the MAL loci genetically demonstrated to be present in CB11. Our results indicate that the cloned maltase structural gene in plasmid pMAL9-26 is linked to MAL6. Since the MAL6 locus has previously been shown to contain a regulatory gene, the MAL6 locus must be a complex locus containing at least two of the factors needed for maltose fermentation: the structural gene for maltase and the maltase regulatory protein. The absence of other fragments which hybridize to the MAL6 -derived probe shows that either MAL2 and MAL4 are not related to MAL6 , or the DNA corresponding to these genes is absent from the MAL6 strain CB11.

scholarly journals Repeated Family of Genes Controlling Maltose Fermentation inSaccharomyces carlsbergensis

1983 ◽  
Vol 3 (5) ◽  
pp. 796-802 ◽  
Author(s):  
Richard B. Needleman ◽  
Corinne Michels
Keyword(s):  
Genetic Analysis ◽  
Structural Gene ◽  
Regulatory Gene ◽  
Regulatory Protein ◽  
Physical Analysis ◽  
Maltose Permease ◽  
Maltose Fermentation ◽  
Complex Locus ◽  
Dna Fragment

Maltose fermentation inSaccharomycesspp. requires the presence of any one of five unlinked genes:MAL1, MAL2, MAL3, MAL4, orMAL6.Although the genes are functionally equivalent, their natures and relationships to each other are not known. At least three proteins are necessary for maltose fermentation: maltase, maltose permease, and a regulatory protein. TheMALgenes may code for one or more of these proteins. Recently a DNA fragment containing a maltase structural gene has been cloned from aMAL6strain, CB11, to produce plasmid pMAL9-26. We have conducted genetic and physical analyses of strain CB11. The genetic analysis has demonstrated the presence of two crypticMALgenes in CB11,MAL1gandMAL3g(linked toMAL1and toMAL3, respectively), in addition to theMAL6locus. The physical analysis, which used a subclone of plasmid pMAL9-26 as a probe, detected threeHindIII genomic fragments with homology to the probe. Each fragment was shown to be linked to one of theMALloci genetically demonstrated to be present in CB11. Our results indicate that the cloned maltase structural gene in plasmid pMAL9-26 is linked toMAL6.Since theMAL6locus has previously been shown to contain a regulatory gene, theMAL6locus must be a complex locus containing at least two of the factors needed for maltose fermentation: the structural gene for maltase and the maltase regulatory protein. The absence of other fragments which hybridize to theMAL6-derived probe shows that eitherMAL2andMAL4are not related toMAL6, or the DNA corresponding to these genes is absent from theMAL6strain CB11.

Identification of a second trans-acting gene controlling maltose fermentation in Saccharomyces carlsbergensis

1986 ◽  
Vol 6 (8) ◽  
pp. 2757-2765
Author(s):  
R A Dubin ◽  
E L Perkins ◽  
R B Needleman ◽  
C A Michels
Keyword(s):  
Structural Gene ◽  
Regulatory Gene ◽  
Constitutive Expression ◽  
Gene Mutations ◽  
Gene Products ◽  
Structural Genes ◽  
Maltose Permease ◽  
Maltose Fermentation ◽  
New Gene

Maltose fermentation in Saccharomyces spp. requires the presence of a dominant MAL locus. The MAL6 locus has been cloned and shown to encode the structural genes for maltose permease (MAL61), maltase (MAL62), and a positively acting regulatory gene (MAL63). Induction of the MAL61 and MAL62 gene products requires the presence of maltose and the MAL63 gene. Mutations within the MAL63 gene produce nonfermenting strains unable to induce the two structural gene products. Reversion of these mal63 nonfermenters to maltose fermenters nearly always leads to the constitutive expression of maltase and maltose permease, and constitutivity is always linked to MAL6. We demonstrated that for one such revertant, strain C2, constitutivity did not require the MAL63 gene, since deletion disruption of this gene did not affect the constitutive expression of the structural genes. In addition, constitutivity was trans acting. Deletion disruption of the MAL6-linked structural genes for maltase and maltose permease in this strain did not affect the constitutive expression of a second, unlinked maltase structural gene. We isolated new maltose-fermenting revertants of a nonfermenting strain which carried a deletion disruption of the MAL63 gene. All 16 revertants isolated expressed maltase constitutively. In one revertant studied in detail, strain R10, constitutive expression was demonstrated to be linked to MAL6, semidominant, trans acting, and residing outside the MAL63-MAL61-MAL62 genes. From these studies we propose the existence of a second trans-acting regulatory gene at the MAL6 locus. We call this new gene MAL64. We mapped the MAL64 gene 2.3 centimorgans to the left of MAL63. The role of the MAL64 gene product in maltose fermentation is discussed.

scholarly journals Identification of a second trans-acting gene controlling maltose fermentation in Saccharomyces carlsbergensis.

1986 ◽  
Vol 6 (8) ◽  
pp. 2757-2765 ◽  
Author(s):  
R A Dubin ◽  
E L Perkins ◽  
R B Needleman ◽  
C A Michels
Keyword(s):  
Structural Gene ◽  
Regulatory Gene ◽  
Constitutive Expression ◽  
Gene Mutations ◽  
Gene Products ◽  
Structural Genes ◽  
Maltose Permease ◽  
Maltose Fermentation ◽  
New Gene

Maltose fermentation in Saccharomyces spp. requires the presence of a dominant MAL locus. The MAL6 locus has been cloned and shown to encode the structural genes for maltose permease (MAL61), maltase (MAL62), and a positively acting regulatory gene (MAL63). Induction of the MAL61 and MAL62 gene products requires the presence of maltose and the MAL63 gene. Mutations within the MAL63 gene produce nonfermenting strains unable to induce the two structural gene products. Reversion of these mal63 nonfermenters to maltose fermenters nearly always leads to the constitutive expression of maltase and maltose permease, and constitutivity is always linked to MAL6. We demonstrated that for one such revertant, strain C2, constitutivity did not require the MAL63 gene, since deletion disruption of this gene did not affect the constitutive expression of the structural genes. In addition, constitutivity was trans acting. Deletion disruption of the MAL6-linked structural genes for maltase and maltose permease in this strain did not affect the constitutive expression of a second, unlinked maltase structural gene. We isolated new maltose-fermenting revertants of a nonfermenting strain which carried a deletion disruption of the MAL63 gene. All 16 revertants isolated expressed maltase constitutively. In one revertant studied in detail, strain R10, constitutive expression was demonstrated to be linked to MAL6, semidominant, trans acting, and residing outside the MAL63-MAL61-MAL62 genes. From these studies we propose the existence of a second trans-acting regulatory gene at the MAL6 locus. We call this new gene MAL64. We mapped the MAL64 gene 2.3 centimorgans to the left of MAL63. The role of the MAL64 gene product in maltose fermentation is discussed.

scholarly journals Genetic and physical analyses of sister chromatid exchange in yeast meiosis

1991 ◽  
Vol 11 (12) ◽  
pp. 6328-6336
Author(s):  
H Sun ◽  
D Dawson ◽  
J W Szostak
Keyword(s):  
Genetic Analysis ◽  
Gene Conversion ◽  
Mutant Strain ◽  
Sister Chromatid ◽  
Sister Chromatid Exchange ◽  
Physical Analysis ◽  
Single Stranded Dna ◽  
Meiotic Gene ◽  
Dna Fragment ◽  
Yeast Meiosis

We have used nonessential circular minichromosomes to monitor sister chromatid exchange during yeast meiosis. Genetic analysis shows that a 64-kb circular minichromosome undergoes sister chromatid exchange during 40% of meioses. This frequency is not reduced by the presence of a homologous linear minichromosome. Furthermore, sister chromatid exchange can be stimulated by the presence of a 12-kb ARG4 DNA fragment, which contains initiation sites for meiotic gene conversion. Using physical analysis, we have directly identified a product of sister chromatid exchange: a head-to-tail dimer form of a circular minichromosome. This dimer form is absent in a rad50S mutant strain, which is deficient in processing of the ends of meiosis-specific double-stranded breaks into single-stranded DNA tails. Our studies suggest that meiotic sister chromatid exchange is stimulated by the same mechanism as meiotic homolog exchange.

Structural and functional analysis of the MAL1 locus of Saccharomyces cerevisiae

1986 ◽  
Vol 6 (11) ◽  
pp. 3891-3899
Author(s):  
M J Charron ◽  
R A Dubin ◽  
C A Michels
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Functional Analysis ◽  
Maltose Permease ◽  
Positive Regulator ◽  
Maltose Fermentation ◽  
Complex Locus ◽  
Regulator Genes

We describe the isolation of a 22.6-kilobase fragment of DNA containing the MAL1 locus of Saccharomyces cerevisiae. Our results demonstrate that the MAL1 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 MAL11, 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 MAL1 locus and in strains carrying single disruptions in each of the MAL1 genes. The MAL1 and MAL1 loci were found to be highly sequence homologous and conserved throughout the region containing these three genes. The strain used to isolate the MAL1 locus also carried the tightly linked SUC1 gene. The SUC1 gene was found to be located on the same 22.6-kilobase fragment containing the MAL1 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.

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

1986 ◽  
Vol 6 (11) ◽  
pp. 3891-3899 ◽  
Author(s):  
M J Charron ◽  
R A Dubin ◽  
C A Michels
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Functional Analysis ◽  
Maltose Permease ◽  
Positive Regulator ◽  
Maltose Fermentation ◽  
Complex Locus ◽  
Regulator Genes

We describe the isolation of a 22.6-kilobase fragment of DNA containing the MAL1 locus of Saccharomyces cerevisiae. Our results demonstrate that the MAL1 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 MAL11, 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 MAL1 locus and in strains carrying single disruptions in each of the MAL1 genes. The MAL1 and MAL1 loci were found to be highly sequence homologous and conserved throughout the region containing these three genes. The strain used to isolate the MAL1 locus also carried the tightly linked SUC1 gene. The SUC1 gene was found to be located on the same 22.6-kilobase fragment containing the MAL1 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.

scholarly journals Genetic and physical analyses of sister chromatid exchange in yeast meiosis.

1991 ◽  
Vol 11 (12) ◽  
pp. 6328-6336 ◽  
Author(s):  
H Sun ◽  
D Dawson ◽  
J W Szostak
Keyword(s):  
Genetic Analysis ◽  
Gene Conversion ◽  
Mutant Strain ◽  
Sister Chromatid ◽  
Sister Chromatid Exchange ◽  
Physical Analysis ◽  
Single Stranded Dna ◽  
Meiotic Gene ◽  
Dna Fragment ◽  
Yeast Meiosis

We have used nonessential circular minichromosomes to monitor sister chromatid exchange during yeast meiosis. Genetic analysis shows that a 64-kb circular minichromosome undergoes sister chromatid exchange during 40% of meioses. This frequency is not reduced by the presence of a homologous linear minichromosome. Furthermore, sister chromatid exchange can be stimulated by the presence of a 12-kb ARG4 DNA fragment, which contains initiation sites for meiotic gene conversion. Using physical analysis, we have directly identified a product of sister chromatid exchange: a head-to-tail dimer form of a circular minichromosome. This dimer form is absent in a rad50S mutant strain, which is deficient in processing of the ends of meiosis-specific double-stranded breaks into single-stranded DNA tails. Our studies suggest that meiotic sister chromatid exchange is stimulated by the same mechanism as meiotic homolog exchange.

scholarly journals GENETIC ANALYSIS OF THE GENE FOR N-ACETYLGLUCOSAMINIDASE IN DICTYOSTELIUM DISCOIDEUM

Genetics ◽  
1978 ◽  
Vol 88 (2) ◽  
pp. 277-284 ◽  
Author(s):  
William F Loomis
Keyword(s):  
Linkage Group ◽  
Genetic Analysis ◽  
Dictyostelium Discoideum ◽  
Structural Gene ◽  
Regulatory Protein ◽  
Group Iv ◽  
Single Locus ◽  
Stages Of Development ◽  
Separate Locus

ABSTRACT Three independent mutations affecting N-acetylglucosaminidase in Dictyostelium discoideum were mapped by the parasexual system and found to lie on linkage group IV. These mutations as well as two others were found to be recessive and noncomplementing in heterozygous diploids. Thus they all appear to affect the nagA locus. Since two of the mutations give rise to thermolabile enzyme, this defines the structural gene for N-acetylglucosaminidase. The enzyme is a homodimer of a 68,000 dalton subunit and thus would be expected to be determined by a single locus. The expression of this gene is regulated by the stages of development; however, it should be mentioned that none of the mutations fell in a separate locus that might determine a specific positive regulatory protein.

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

1988 ◽  
Vol 8 (3) ◽  
pp. 1027-1035
Author(s):  
R A Dubin ◽  
M J Charron ◽  
S R Haut ◽  
R B Needleman ◽  
C A Michels
Keyword(s):  
Constitutive Expression ◽  
Positive Function ◽  
Gene Products ◽  
Wild Type ◽  
Transcript Accumulation ◽  
Saccharomyces Carlsbergensis ◽  
Maltose Permease ◽  
Maltose Fermentation ◽  
Mutational Activation ◽  
Type Strains

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.

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

1988 ◽  
Vol 8 (3) ◽  
pp. 1027-1035 ◽  
Author(s):  
R A Dubin ◽  
M J Charron ◽  
S R Haut ◽  
R B Needleman ◽  
C A Michels
Keyword(s):  
Constitutive Expression ◽  
Positive Function ◽  
Gene Products ◽  
Wild Type ◽  
Transcript Accumulation ◽  
Saccharomyces Carlsbergensis ◽  
Maltose Permease ◽  
Maltose Fermentation ◽  
Mutational Activation ◽  
Type Strains

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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