Abstract
A sensitive assay using biotinylated ubiquitin revealed extensive ubiquitination of the large subunit of RNA polymerase II during incubations of transcription reactions in vitro. Phosphorylation of the repetitive carboxyl-terminal domain of the large subunit was a signal for ubiquitination. Specific inhibitors of cyclin-dependent kinase (cdk)-type kinases suppress the ubiquitination reaction. These kinases are components of transcription factors and have been shown to phosphorylate the carboxyl-terminal domain. In both regulation of transcription and DNA repair, phosphorylation of the repetitive carboxyl-terminal domain by kinases might signal degradation of the polymerase.
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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Amanitins / pharmacology
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Cell Nucleus / metabolism
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Cyclin-Dependent Kinases / antagonists & inhibitors
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Cysteine Endopeptidases / metabolism
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Enzyme Inhibitors / pharmacology
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Glutathione Transferase / metabolism
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HeLa Cells
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Humans
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Kinetics
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Macromolecular Substances
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Multienzyme Complexes / metabolism
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Peptide Fragments / chemistry
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Peptide Fragments / metabolism
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Phosphorylation
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Proteasome Endopeptidase Complex
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RNA Polymerase II / chemistry*
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RNA Polymerase II / metabolism*
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / metabolism
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Templates, Genetic
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Transcription, Genetic
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Ubiquitins / metabolism*
Substances
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Amanitins
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Enzyme Inhibitors
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Macromolecular Substances
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Multienzyme Complexes
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Peptide Fragments
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Recombinant Fusion Proteins
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Ubiquitins
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Glutathione Transferase
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Cyclin-Dependent Kinases
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RNA Polymerase II
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Cysteine Endopeptidases
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Proteasome Endopeptidase Complex