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|標題:||RNA polymerase II is released from the DNA template during transcription-coupled repair in mammalian cells||作者:||Chiou, Yi-Ying
Selby, Christopher P
|關鍵字:||DNA damage;DNA repair;RNA polymerase II;gene transcription;nucleotide excision repair;DNA;DNA Damage;Fibroblasts;Humans;RNA Polymerase II;Templates, Genetic;Ultraviolet Rays;DNA Repair;Transcription, Genetic||Project:||The Journal of biological chemistry, Volume 293, Issue 7, Page(s) 2476-2486.||摘要:||
In mammalian cells, bulky DNA adducts located in the template but not the coding strand of genes block elongation by RNA polymerase II (RNAPII). The blocked RNAPII targets these transcription-blocking adducts to undergo more rapid excision repair than adducts located elsewhere in the genome. In excision repair, coupled incisions are made in the damaged DNA strand on both sides of the adduct. The fate of RNAPII in the course of this transcription-coupled repair (TCR) pathway is unclear. To address the fate of RNAPII, we used methods that control transcription to initiate a discrete "wave" of elongation complexes. Analyzing genome-wide transcription and repair by next-generation sequencing, we identified locations of elongation complexes and transcription-repair coupling events in genes throughout the genome. Using UV-exposed human skin fibroblasts, we found that, at the dose used, a single wave of elongation complexes was blocked within the first 25 kb of genes. TCR occurred where the elongation complexes were blocked, and repair was associated with the dissociation of these complexes. These results indicate that individual elongation complexes do not engage in multiple rounds of TCR with successive lesions. Our results are consistent with a model in which RNAPII is dissociated after the dual incision of the transcription-blocking lesion, perhaps by Cockayne syndrome group B translocase, or during the synthesis of a repair patch.
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