Interaction of GAL4 and GAL80 gene regulatory proteins in vitro.
The GAL80 protein of Saccharomyces cerevisiae, synthesized in vitro, bound tightly to GAL4 protein and to a GAL4 protein-upstream activation sequence DNA complex, as shown by (i) coimmunoprecipitation of GAL4 and GAL80 proteins with anti-GAL4 antiserum, (ii) an electrophoretic mobility shift of a GAL4 protein-upstream activation sequence DNA complex upon the addition of GAL80 protein, and (iii) GAL4-dependent binding of GAL80 protein to upstream activation sequence DNA immobilized on Sepharose beads. Anti-GAL4 antisera were raised against a GAL4-URA3 fusion protein, which could be purified to homogeneity in a single step with the use of an affinity chromatographic procedure for the URA3 gene product.
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Department of Cell Biology, Stanford University School of Medicine, California 94305.
Chasman
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Buchman
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Kornberg
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D004271:DNA, Fungal / Q000235:genetics; D004268:DNA-Binding Proteins / Q000502:physiology*; D005656:Fungal Proteins / Q000502:physiology; D005690:Galactose / Q000502:physiology*; D005786:Gene Expression Regulation; D007158:Immunologic Techniques; D046911:Macromolecular Substances; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000276:immunology; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014157:Transcription Factors / Q000502:physiology*
D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D046911:Macromolecular Substances; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D005690:Galactose