Abstract
Recognition of the TATA box by the TATA-binding protein (TBP) is a highly regulated step in RNA polymerase II-dependent transcription. Several proteins have been proposed to regulate TBP activity, yet the TBP domains responsive to all these regulators have not been defined. Here we describe a new class of TBP mutants that increase transcription from core promoters in vivo. The majority of these mutations alter amino acids that cluster tightly on the TBP surface, defining a new TBP regulatory domain. The mutant TBP proteins are defective for binding the transcriptional regulator yNC2, are resistant to inhibition by yNC2 in vitro and exhibit allele-specific genetic interactions with yNC2. These results provide strong biochemical and genetic evidence that TBP is directly repressed in vivo, and define a new TBP domain important for transcriptional regulation.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Adenosine Triphosphatases
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Alleles
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DNA Helicases / metabolism
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DNA-Binding Proteins / chemistry*
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DNA-Binding Proteins / genetics
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Gene Expression Regulation, Fungal
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Genetic Complementation Test
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Models, Molecular
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Mutagenesis
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Phenotype
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Phosphoproteins / chemistry
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Phosphoproteins / metabolism
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Plasmids
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Promoter Regions, Genetic
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Protein Binding
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Protein Structure, Tertiary
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Repressor Proteins / metabolism
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Saccharomyces cerevisiae / chemistry
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Saccharomyces cerevisiae Proteins*
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TATA-Binding Protein Associated Factors*
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TATA-Box Binding Protein
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Transcription Factor TFIIA
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Transcription Factors / chemistry*
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Transcription Factors / genetics
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Transcription Factors / metabolism
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Transcription, Genetic
Substances
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BUR6 protein, S cerevisiae
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DNA-Binding Proteins
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Phosphoproteins
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Repressor Proteins
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Saccharomyces cerevisiae Proteins
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TATA-Binding Protein Associated Factors
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TATA-Box Binding Protein
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Transcription Factor TFIIA
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Transcription Factors
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down-regulator of transcription 1
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Adenosine Triphosphatases
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MOT1 protein, S cerevisiae
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DNA Helicases