The structural genes coding for the two kinds of subunits of phosphofructokinase in yeast have been cloned previously (Heinisch 1986). The coding regions were defined by S1-mapping. They were disrupted in vitro by insertion of a LEU2-marker. These constructions were then used for substitution of the respective chromosomal copies. That the disruption of the PFK-genes had in fact occurred was confirmed by Southern blot analysis. Furthermore, in Northern blots shorter transcripts were detected in the respective disruption mutants. Using polyclonal antibodies the alpha-subunits were not detectable in pfk1-disruptions whereas the beta-subunits were undetectable in pfk2-disruptions. Physiological characterization showed that the single disruption mutants still fermented glucose to ethanol and CO2. They accumulated fructose-6-phosphate and glucose-6-phosphate over wild type levels and showed decreased levels of fructose-1,6-bisphosphate. In addition an accumulation of sedoheptulose-7-phosphate was observed, a metabolite not detectable in wild type cells. A haploid yeast strain containing both disrupted copies of the PFK-genes is not capable of growing on rich medium containing 2% glucose. The accumulation of glucose-6-phosphate, fructose-6-phosphate and sedoheptulose-7-phosphate is much more pronounced in such mutants, whereas the fructose-1,6-bisphosphate concentration decreases below the level of detection.
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Evidence ID | Analyze ID | Gene/Complex | Systematic Name/Complex Accession | Qualifier | Gene Ontology Term ID | Gene Ontology Term | Aspect | Annotation Extension | Evidence | Method | Source | Assigned On | Reference |
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Evidence ID | Analyze ID | Gene | Gene Systematic Name | Phenotype | Experiment Type | Experiment Type Category | Mutant Information | Strain Background | Chemical | Details | Reference |
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Evidence ID | Analyze ID | Gene | Gene Systematic Name | Disease Ontology Term | Disease Ontology Term ID | Qualifier | Evidence | Method | Source | Assigned On | Reference |
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Evidence ID | Analyze ID | Regulator | Regulator Systematic Name | Target | Target Systematic Name | Direction | Regulation of | Happens During | Regulator Type | Direction | Regulation Of | Happens During | Method | Evidence | Strain Background | Reference |
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Site | Modification | Modifier | Source | Reference |
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Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Allele | Assay | Annotation | Action | Phenotype | SGA score | P-value | Source | Reference | Note |
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Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Assay | Annotation | Action | Modification | Source | Reference | Note |
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Complement ID | Locus ID | Gene | Species | Gene ID | Strain background | Direction | Details | Source | Reference |
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Evidence ID | Analyze ID | Dataset | Description | Keywords | Number of Conditions | Reference |
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