Abstract
Mig1p is a zinc finger protein required for repression of glucose-regulated genes in budding yeast. On removal of medium glucose, gene repression is relieved via a mechanism that requires the SNF1 protein kinase complex. We show that Mig1p expressed as a glutathione-S-transferase fusion in bacteria is readily phosphorylated by the SNF1 kinase in vitro. Four phosphorylation sites were identified, i.e. Ser-222, Ser-278, Ser-311 and Ser-381. The latter three are exact matches to the recognition motif we previously defined for SNF1 and lie within regions shown to be required for SNF1-dependent derepression and nuclear-to-cytoplasmic translocation.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Aspartic Acid / genetics
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism*
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Enzyme Repression
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Gene Expression Regulation, Fungal
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Glucose / metabolism
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Glutamic Acid / genetics
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Glutathione Transferase / genetics
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Kluyveromyces / enzymology
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Kluyveromyces / genetics
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Mutagenesis, Site-Directed
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Phosphorylation
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Protein Serine-Threonine Kinases / genetics
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Protein Serine-Threonine Kinases / metabolism*
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Recombinant Fusion Proteins / metabolism
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Regulatory Sequences, Nucleic Acid
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Repressor Proteins / genetics
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Repressor Proteins / metabolism*
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Saccharomyces cerevisiae / enzymology*
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Saccharomyces cerevisiae / genetics
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Saccharomyces cerevisiae Proteins
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Serine / genetics
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Substrate Specificity
Substances
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DNA-Binding Proteins
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MIG1 protein, S cerevisiae
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Recombinant Fusion Proteins
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Repressor Proteins
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Saccharomyces cerevisiae Proteins
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Aspartic Acid
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Glutamic Acid
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Serine
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Glutathione Transferase
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SNF1-related protein kinases
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Protein Serine-Threonine Kinases
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Glucose