Abstract
The FACT (facilitates chromatin transcription) complex is required for transcript elongation through nucleosomes by RNA polymerase II (Pol II) in vitro. Here, we show that FACT facilitates Pol II-driven transcription by destabilizing nucleosomal structure so that one histone H2A-H2B dimer is removed during enzyme passage. We also demonstrate that FACT possesses intrinsic histone chaperone activity and can deposit core histones onto DNA. Importantly, FACT activity requires both of its constituent subunits and is dependent on the highly acidic C terminus of its larger subunit, Spt16. These findings define the mechanism by which Pol II can transcribe through chromatin without disrupting its epigenetic status.
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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Amino Acid Sequence
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Animals
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Cell Cycle Proteins / chemistry
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Cell Cycle Proteins / metabolism
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Cell Line
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DNA / metabolism
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DNA-Binding Proteins / chemistry
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DNA-Binding Proteins / metabolism
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Dimerization
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HeLa Cells
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High Mobility Group Proteins / chemistry
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High Mobility Group Proteins / metabolism
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Histones / metabolism
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Humans
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Models, Genetic
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Molecular Chaperones / chemistry
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Molecular Chaperones / metabolism
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Molecular Sequence Data
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Nucleosomes / metabolism*
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Protein Binding
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Protein Subunits
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RNA Polymerase II / metabolism*
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Recombinant Proteins / chemistry
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Recombinant Proteins / metabolism
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Templates, Genetic
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Transcription Factors / chemistry
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Transcription Factors / metabolism
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Transcription, Genetic*
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Transcriptional Elongation Factors / chemistry
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Transcriptional Elongation Factors / metabolism*
Substances
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Cell Cycle Proteins
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DNA-Binding Proteins
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High Mobility Group Proteins
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Histones
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Molecular Chaperones
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Nucleosomes
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Protein Subunits
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Recombinant Proteins
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SSRP1 protein, human
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SUPT16H protein, human
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Transcription Factors
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Transcriptional Elongation Factors
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DNA
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RNA Polymerase II