ミウラ　ミチ   Michi Miura   三浦　未知    所属   川崎医科大学  医学部 基礎医学　微生物学    職種   助教
論文種別	原著
言語種別	英語
査読の有無	査読あり
表題	CUT&RUN detects distinct DNA footprints of RNA polymerase II near the transcription start sites.
掲載誌名	正式名：Chromosome Research 略　 称：Chromosome Res ISSNコード：15736849/09673849
掲載区分	国外
巻・号・頁	28(3-4),pp.381-393
著者・共著者	Miura Michi, Chen Honglin
担当区分	筆頭著者,責任著者
発行年月	2020/12
概要	CUT&RUN is a powerful tool to study protein-DNA interactions in vivo. DNA fragments cleaved by the targeted micrococcal nuclease identify the footprints of DNA-binding proteins on the chromatin. We performed CUT&RUN on human lung carcinoma cell line A549 maintained in a multi-well cell culture plate to profile RNA polymerase II. Long (> 270 bp) DNA fragments released by CUT&RUN corresponded to the bimodal peak around the transcription start sites, as previously seen with chromatin immunoprecipitation. However, we found that short (< 120 bp) fragments identify a well-defined peak localised at the transcription start sites. This distinct DNA footprint of short fragments, which constituted only about 5% of the total reads, suggests the transient positioning of RNA polymerase II before promoter-proximal pausing, which has not been detected in the physiological settings by standard chromatin immunoprecipitation. We showed that the positioning of the large-size-class DNA footprints around the short-fragment peak was associated with the directionality of transcription, demonstrating the biological significance of distinct CUT&RUN footprints of RNA polymerase II.
DOI	10.1007/s10577-020-09643-0
PMID	33070289