Abstract
The TATA-binding protein TBP has been recently recognized as a general class III transcription factor. Using the gel shift assay to monitor initiation complex assembly on a yeast tRNA gene, we show that TBP is required for the TFIIIC-dependent assembly of TFIIIB. TFIIIB depleted of TBP by a simple chromatographic step does not bind stably to the TFIIIC-tDNA complex. Addition of yeast or human recombinant TBP allows the formation of a TFIIIB-TBP-TFIIIC-tDNA complex. The presence of TBP in the complex was inferred from the effect of anti-TBP antibodies and from the different migration properties of TFIIIB-TBP-tDNA complexes formed with yeast or human TBP.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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DNA, Fungal / genetics
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DNA, Fungal / metabolism
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DNA-Binding Proteins / metabolism*
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Genes, Fungal
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Humans
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RNA, Transfer / genetics*
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Recombinant Proteins / metabolism
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Saccharomyces cerevisiae / genetics
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Saccharomyces cerevisiae / metabolism*
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TATA-Box Binding Protein
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Transcription Factor TFIIIB
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Transcription Factors / biosynthesis*
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Transcription Factors / genetics
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Transcription Factors / isolation & purification
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Transcription Factors / metabolism*
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Transcription Factors, TFIII*
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Transcription, Genetic
Substances
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DNA, Fungal
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DNA-Binding Proteins
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Recombinant Proteins
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TATA-Box Binding Protein
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Transcription Factor TFIIIB
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Transcription Factors
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Transcription Factors, TFIII
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transcription factor TFIIIC
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RNA, Transfer