Isolation and characterization of Saccharomyces cerevisiae glycolytic pathway mutants

Lam, Keng-Bon; Marmur, Julius

doi:10.1128/jb.130.2.746-749.1977

Cited by 46 publications

(16 citation statements)

References 11 publications

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“…The transcriptome analysis performed in this study indicates that the severely impaired growth phenotype of the Δgpm1 strain is associated with a global stress response.As it has been previously shown [3,4], a Δgpm1 strain is incapable of growing on glucose as the sole carbon source but growth can be restored in presence of a mixture of glycerol and ethanol (2-3% v/v). Previous studies [1] have suggested that ethanol is respired whereas glycerol is needed to feed gluconeogenesis.…”

Section: Discussionmentioning

confidence: 86%

“…It was previously shown that a S. cerevisiae strain deleted in the glycolytic gene GPM1 (encoding phosphoglycerate mutase) is not able to use glucose as sole carbon source but has the capability to grow on a mixture of ethanol and glycerol [4]. Here we studied the physiological response of a Δgpm1 strain in batch cultivations in minimal media supplemented with glycerol (3% v/v) and ethanol 2.5% (v/v) as carbon sources.…”

Section: Physiological Characterization Of the Deletion Strain δ δGpmmentioning

confidence: 99%

“…In the yeast Saccharomyces cerevisiae, this enzyme is a tetramer composed of four identical subunits with a molecular weight of 28 000 Da each [1]. The gene was firstly isolated through functional complementation [2] of mutants classified among glycolytic mutants whose growth was inhibited by glucose [3,4]. GPM1 is a constitutively expressed glycolytic gene and its promoter contains a variety of elements mediating transcriptional control [5].…”

Section: Introductionmentioning

confidence: 99%

“…Nevertheless, the role of Gpm1 in the cell wall remains largely unknown. Mutation of GPM1 in the yeast genome leads to viable cells only when glycerol and ethanol are provided together as carbon source, whilst growth of the mutant is inhibited in the presence of glucose or ethanol or glycerol alone [4]. It has been suggested that glycerol is needed to feed gluconeogenesis whereas Gibbs free energy is provided through respiration of ethanol.…”

Section: Introductionmentioning

confidence: 99%

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Phosphoglycerate mutase knock‐out mutant Saccharomyces cerevisiae: Physiological investigation and transcriptome analysis

Papini

Nookaew

Scalcinati

et al. 2010

Biotechnology Journal

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The yeast Saccharomyces cerevisiae is able to adapt its metabolism to grow on different carbon sources and to shift to non‐fermentative growth on C2 or C3 carbon sources (ethanol, acetate, or glycerol) through the activation of gluconeogenesis. Here, we studied the response to the deletion of the glycolytic and gluconeogenic gene GPM1, encoding for phosphoglycerate mutase. It was previously shown that a S. cerevisiae strain with non‐functional copies of GPM1 can only grow when glycerol and ethanol are both present as carbon sources, whilst addition of glucose was shown to strongly inhibit growth. It was suggested that glycerol is needed to feed gluconeogenesis whilst ethanol is required for respiration. Here, we studied the physiological response of the GPM1 knock‐out mutant through fermentation and transcriptome analysis. Furthermore, we compared the physiological results with those obtained through simulations using a genome‐scale metabolic model, showing that glycerol is only needed in small amounts for growth. Our findings strongly suggest a severely impaired growth ability of the knock‐out mutant, which presents increased transcript levels of genes involved in the pentose phosphate pathway and in the glyoxylate shunt. These results indicate an attempt to compensate for the energy imbalance caused by the deletion of the glycolytic/gluconeogenic gene within the mutant.

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Section: Discussionmentioning

confidence: 86%

Section: Physiological Characterization Of the Deletion Strain δ δGpmmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Phosphoglycerate mutase knock‐out mutant Saccharomyces cerevisiae: Physiological investigation and transcriptome analysis

Papini

Nookaew

Scalcinati

et al. 2010

Biotechnology Journal

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“…Several yeast glycolytic mutants have been isolated recently (3,7,14,15,23), but only mutants lacking hexokinases (16), phosphoglucose isomerase (18), or pyruvate kinase (20,23) were characterized in detail. The common feature of all of those mutants was that growth was inhibited by glucose.…”

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confidence: 99%

Physiological Effects of Seven Different Blocks in Glycolysis inSaccharomyces cerevisiae

1979

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Saccharomyces cerevisiae mutants unable to grow and ferment glucose have been isolated. Of 45 clones isolated, 25 had single enzyme defects of one of the following activities: phosphoglucose isomerase (pgi), phosphofructokinase (pfk), triosephosphate isomerase (tpi), phosphoglycerate kinase (pgk), phosphoglyceromutase (pgm), and pyruvate kinase (pyk). Phosphofructokinase activities in crude extracts of the pfk mutant were only 2% of the wild-type level. However, nornal growth on glucose medium and normal fermentation of glucose suggested either that the mutant enzyme was considerably more active in vivo or, alternatively, that 2% residual activity was sufficient for normal glycolysis. All other mutants were moderately to strongly inhibited by glucose. Unusually high concentrations of glycolytic metabolites were observed before the reaction catalyzed by the enzyme which was absent in a given mutant strain when incubated on glucose. This confirmed at the cellular level the location of the defect as determined by enzyme assays. With adh (lacks all three alcohol dehydrogenase isozymes) and pgk mutants, accumulation of the typical levels of hexosephosphates was prevented when respiration was blocked with antimycin A. A typical feature of all glycolytic mutants described here was the rapid depletion of the intracellular adenosine 5'-triphosphate pool after transfer to glucose medium. No correlation of low or high levels of fructose-1,6-bisphosphate with the degree of catabolite repression and inactivation could be found. This observation does not support the concept that hexose metabolites are directly involved in these regulatory mechanisms in yeast.

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Investigation of two yeast genes encoding putative isoenzymes of phosphoglycerate mutase

Heinisch

Müller

Schlüter

et al. 1998

Yeast

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Our previous data indicated that GPM1 encodes the only functional phosphoglycerate mutase in yeast. However, in the course of the yeast genome sequencing project, two homologous sequences, designated GPM2 and GPM3, were detected. They have been further investigated in this work. Key residues in the deduced amino acid sequence, shown to be involved in catalysis for Gpm1 (i.e. His8, Arg59, His181) are conserved in both enzymes. Overexpression of the genes under control of their own promoters in a gpm1 deletion mutant did not complement for any of the phenotypes. This could in part be attributed to a lack of expression due to their weak promoters. Higher level expression under the control of the yeast PFK2 promoter partially complemented the gpm1 defects, without restoring detectable enzymatic activity. Nevertheless, deletion of either GPM2 or GPM3, or the two deletions in concert, did not produce any obvious lesions for growth on a variety of different carbon sources, nor did they change the levels of key intermediary metabolites. We conclude that both genes evolved from duplication events and that they probably constitute non‐functional homologues in yeast.

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Isolation and characterization of Saccharomyces cerevisiae glycolytic pathway mutants

Cited by 46 publications

References 11 publications

Phosphoglycerate mutase knock‐out mutant Saccharomyces cerevisiae: Physiological investigation and transcriptome analysis

Phosphoglycerate mutase knock‐out mutant Saccharomyces cerevisiae: Physiological investigation and transcriptome analysis

Physiological Effects of Seven Different Blocks in Glycolysis inSaccharomyces cerevisiae

Investigation of two yeast genes encoding putative isoenzymes of phosphoglycerate mutase

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