The transcription-repair coupling factor CSA is required for efficient repair only during the elongation stages of RNA polymerase II transcription.
The known nucleotide excision repair (NER) defects of xeroderma pigmentosum (XP) and Cockayne syndrome (CS) cells can be exploited to analyze mechanisms of repair of UV-induced cyclobutane pyrimidine dimers (CPDs) at nucleotide (nt.) resolution. The two gene products of the CS complementation groups (CSA and CSB) have been implicated in the preferential repair of the transcribed strand of human genes. We had previously described very efficient repair of CPDs at sequences near the transcription initiation site of the human JUN gene in normal fibroblasts. Here, we have analyzed repair in a CSA fibroblast strain. CSA cells exhibited rapid repair near the transcription initiation site (positions -45 to +15) but were deficient in repair of sequences on the transcribed strand beginning around nt. +20. There was also no strand-selective repair of sequences further downstream of the start site (+260 to +450). The results suggest that the transcription-repair coupling factor (TRCF) CSA is required for efficient repair only during the elongation stages of RNA polymerase II transcription. We also discuss possible mechanisms of differential repair observed near the transcription initiation site in XP and CS cells and conclude that these in vivo repair data support some recent models obtained from nucleotide excision repair experiments in vitro.
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Beckman Research Institute of the City of Hope, Department of Biology, Duarte, CA 91010, USA.
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D002460:Cell Line; D003057:Cockayne Syndrome / Q000235:genetics; D018387:Codon, Initiator; D004247:DNA / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D004260:DNA Repair / Q000502:physiology* / Q000528:radiation effects; D045643:DNA Repair Enzymes; D005347:Fibroblasts / Q000378:metabolism / Q000528:radiation effects; D005786:Gene Expression Regulation / Q000528:radiation effects; D016758:Genes, jun / Q000528:radiation effects; D006801:Humans; D010441:Peptide Chain Elongation, Translational / Q000235:genetics*; D011506:Proteins / Q000502:physiology*; D012319:RNA Polymerase II / Q000235:genetics*; D014157:Transcription Factors; D014158:Transcription, Genetic / Q000502:physiology* / Q000528:radiation effects; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum / Q000235:genetics
D018387:Codon, Initiator; C095472:ERCC8 protein, human; D011506:Proteins; D014157:Transcription Factors; D004247:DNA; D012319:RNA Polymerase II; D045643:DNA Repair Enzymes