Abstract
The assembly of activated RNA polymerase II (pol II) transcription complexes has been investigated by assaying whether pre-assembly of intermediate complexes reduces the extended time required for start-site melting. The results show that a closed complex requiring factors IIA, IID, and the acidic activator GAL4-AH forms in a rate-limiting step. This directs the templates into a productive assembly pathway. Factor TFIIB is then added rapidly, affording further protection against diversion into nonproductive pathways. These events are followed by a series of rapid steps in which the remaining general factors are assembled onto the template, which is then melted using the energy of ATP hydrolysis.
Publication types
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Research Support, U.S. Gov't, Non-P.H.S.
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Base Sequence
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Cell Extracts
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DNA Polymerase II / genetics*
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DNA Polymerase II / metabolism
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DNA-Binding Proteins
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Fungal Proteins / genetics*
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Fungal Proteins / metabolism
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Gene Expression Regulation / genetics
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HeLa Cells
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Humans
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Macromolecular Substances
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Molecular Sequence Data
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Plasmids / genetics
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Potassium Permanganate
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Saccharomyces cerevisiae Proteins*
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Transcription Factor TFIIA
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Transcription Factor TFIID
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Transcription Factors / genetics*
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Transcription Factors / metabolism
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Transcription, Genetic / genetics*
Substances
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Cell Extracts
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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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Macromolecular Substances
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Saccharomyces cerevisiae Proteins
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Transcription Factor TFIIA
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Transcription Factor TFIID
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Transcription Factors
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Potassium Permanganate
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DNA Polymerase II