Abstract
Cockayne's syndrome (CS) is a disease characterized by developmental and growth defects, sunlight sensitivity, and a defect in transcription-coupled nucleotide excision repair. The two principle proteins involved in CS, CSA and CSB/ERCC6, have been hypothesized to bind RNA polymerase II (Pol II) and link transcription to DNA repair. We have tested CSA and CSB in assays designed to determine their role in transcription-coupled repair. Using a unique oligo(dC)-tailed DNA template, we provide biochemical evidence that CSB/ERCC6 interacts with Pol II molecules engaged in ternary complexes containing DNA and nascent RNA. CSB is a DNA-activated ATPase, and hydrolysis of the ATP beta-gamma phosphoanhydride bond is required for the formation of a stable Pol II-CSB-DNA-RNA complex. Unlike CSB, CSA does not directly bind Pol II.
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 Triphosphatases / metabolism
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Base Sequence
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Cockayne Syndrome / genetics*
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Cockayne Syndrome / metabolism*
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DNA Helicases / genetics*
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DNA Helicases / metabolism*
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DNA Probes / genetics
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DNA Repair / genetics
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DNA Repair Enzymes
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Humans
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In Vitro Techniques
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Molecular Sequence Data
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Mutation
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Poly-ADP-Ribose Binding Proteins
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Protein Binding
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Proteins / genetics
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Proteins / metabolism
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RNA Polymerase II / metabolism*
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Transcription Factors
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Transcription, Genetic
Substances
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DNA Probes
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ERCC8 protein, human
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Poly-ADP-Ribose Binding Proteins
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Proteins
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Recombinant Proteins
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Transcription Factors
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RNA Polymerase II
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Adenosine Triphosphatases
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DNA Helicases
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ERCC6 protein, human
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DNA Repair Enzymes