Abstract
A genetic method, the two-hybrid system, was used to identify four genes encoding proteins that interact with the SNF1 protein kinase from yeast. One of the genes, SIP1, was independently isolated as a multicopy suppressor of defects caused by reduced SNF1 kinase activity, and genetic evidence supports its function in the SNF1 pathway. The SIP1 protein co-immunoprecipitated with SNF1 and was phosphorylated in vitro. Thus, the two-hybrid system, which is applicable to any cloned gene, can be used to detect physical interactions between protein kinases and functionally related substrate proteins.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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AMP-Activated Protein Kinases
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Amino Acid Sequence
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Base Sequence
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DNA-Binding Proteins / metabolism
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Fungal Proteins / genetics*
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Fungal Proteins / metabolism*
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Genes, Fungal
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Molecular Sequence Data
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Mutagenesis
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Phosphorylation
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Plasmids
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Protein Kinases / metabolism*
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Protein Serine-Threonine Kinases*
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Recombinant Fusion Proteins / metabolism*
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Restriction Mapping
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Saccharomyces cerevisiae / enzymology*
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Saccharomyces cerevisiae Proteins*
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Substrate Specificity
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Transcription Factors*
Substances
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DNA-Binding Proteins
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Fungal Proteins
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GAL4 protein, S cerevisiae
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Recombinant Fusion Proteins
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Saccharomyces cerevisiae Proteins
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Transcription Factors
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Protein Kinases
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SNF1-related protein kinases
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Protein Serine-Threonine Kinases
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SIP1 protein, S cerevisiae
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AMP-Activated Protein Kinases