Abstract
The yeast SNF2 (SWI2) protein functions with SNF5, SNF6, SWI1, and SWI3 in the transcriptional activation of many differently regulated genes. These proteins appear to facilitate activation by gene-specific regulatory proteins. SNF2 is highly conserved among eukaryotes and defines a family of proteins with similarity to helicases and nucleic acid-dependent NTPases. Here, we present genetic and biochemical evidence that SNF2 has DNA-stimulated ATPase activity. Mutations in the nucleoside triphosphate (NTP)-binding motif and other conserved motifs impair SNF2 function. Swapping experiments with another member of this family indicate that the helicase-related domains are functionally interchangeable. Finally, bacterially expressed SNF2 protein has ATPase activity that is stimulated by double-stranded DNA, and mutation of the NTP-binding site abolishes this activity. Deletion analysis shows that the helicase-like region of SNF2 is necessary, but not sufficient, for transcriptional activation.
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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Adenosine Triphosphatases / metabolism
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Adenosine Triphosphate / metabolism*
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Base Sequence
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Binding Sites / genetics
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Cell Cycle Proteins*
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Chromatin / metabolism
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Conserved Sequence
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DNA Helicases / genetics
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DNA Mutational Analysis
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DNA, Fungal / analysis
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DNA-Binding Proteins / genetics*
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DNA-Binding Proteins / physiology
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Fungal Proteins / genetics*
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Hydrolysis
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Molecular Sequence Data
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Mutagenesis, Site-Directed
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Nuclear Proteins*
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Oligonucleotide Probes
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Protein Binding
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Recombinant Fusion Proteins / physiology
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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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Transcription Factors / genetics*
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Transcription Factors / physiology
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Transcription, Genetic*
Substances
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Cell Cycle Proteins
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Chromatin
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DNA, Fungal
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DNA-Binding Proteins
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Fungal Proteins
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Nuclear Proteins
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Oligonucleotide Probes
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Recombinant Fusion Proteins
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Saccharomyces cerevisiae Proteins
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Transcription Factors
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Adenosine Triphosphate
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Adenosine Triphosphatases
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SNF2 protein, S cerevisiae
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STH1 protein, S cerevisiae
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DNA Helicases