Abstract
TFC5, the unique and essential gene encoding the B" component of the Saccharomyces cerevisiae RNA polymerase III transcription factor (TF)IIIB has been cloned. It encodes a 594-amino acid protein (67,688 Da). Escherichia coli-produced B" has been used to reconstitute entirely recombinant TFIIIB that is fully functional for TFIIIC-directed, as well as TATA box-dependent, DNA binding and transcription. The DNase I footprints of entirely recombinant TFIIIB, composed of B", the 67-kDa Brf, and TATA box-binding protein, and TFIIIB reconstituted with natural B" are indistinguishable. A truncated form of B" lacking 39 N-terminal and 107 C-terminal amino acids is also functional for transcription.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Base Sequence
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Cloning, Molecular
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DNA Footprinting
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DNA, Fungal / metabolism
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DNA-Binding Proteins / metabolism
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Escherichia coli / genetics
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Fungal Proteins / genetics*
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Genes, Fungal*
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Molecular Sequence Data
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Promoter Regions, Genetic
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Protein Binding
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RNA Polymerase III / metabolism*
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RNA, Transfer / genetics
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Recombinant 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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Sequence Analysis
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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 / metabolism
Substances
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BRF1 protein, S cerevisiae
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DNA, Fungal
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DNA-Binding Proteins
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Fungal Proteins
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Recombinant Proteins
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Saccharomyces cerevisiae 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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RNA, Transfer
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RNA Polymerase III