Abstract
High levels of gene transcription by RNA polymerase II depend on high rates of transcription initiation and reinitiation. Initiation requires recruitment of the complete transcription machinery to a promoter, a process facilitated by activators and chromatin remodelling factors. Reinitiation probably occurs through a different pathway. After initiation, a subset of the transcription machinery remains at the promoter, forming a platform for assembly of a second transcription complex. Here we describe the isolation of a reinitiation intermediate that includes transcription factors TFIID, TFIIA, TFIIH, TFIIE and Mediator. This intermediate can act as a scaffold for formation of a functional reinitiation complex. Formation of this scaffold is dependent on ATP and TFIIH. The scaffold is stabilized in the presence of the activator Gal4-VP16, but not Gal4-AH, suggesting a new role for some activators and Mediator in promoting high levels of transcription.
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 Triphosphate / metabolism
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DNA, Fungal / metabolism
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DNA-Binding Proteins
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Fungal Proteins / metabolism
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Macromolecular Substances
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Promoter Regions, Genetic
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RNA Polymerase II / chemistry
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RNA Polymerase II / metabolism
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RNA, Fungal / metabolism
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Saccharomyces cerevisiae Proteins*
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TATA-Binding Protein Associated Factors*
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Trans-Activators / metabolism
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Transcription Factor TFIIA
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Transcription Factor TFIID*
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Transcription Factor TFIIH
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Transcription Factors / chemistry
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Transcription Factors / metabolism*
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Transcription Factors, TFII*
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Transcription, Genetic*
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Yeasts / genetics
Substances
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DNA, Fungal
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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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Gal-VP16
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Macromolecular Substances
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RNA, Fungal
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Saccharomyces cerevisiae Proteins
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TAF6 protein, S cerevisiae
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TATA-Binding Protein Associated Factors
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Trans-Activators
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Transcription Factor TFIIA
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Transcription Factor TFIID
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Transcription Factors
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Transcription Factors, TFII
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transcription factor TFIIE
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Transcription Factor TFIIH
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Adenosine Triphosphate
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RNA Polymerase II