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脱氧核糖核酸酶I足迹法显示非洲爪蟾核糖体RNA基因启动子中有三个受保护区域。

DNase I footprinting shows three protected regions in the promoter of the rRNA genes of Xenopus laevis.

作者信息

Dunaway M, Reeder R H

出版信息

Mol Cell Biol. 1985 Feb;5(2):313-9. doi: 10.1128/mcb.5.2.313-319.1985.

Abstract

Extracts prepared from Xenopus laevis oocytes contain a protein(s) which specifically protects three discrete regions of the RNA polymerase I promoter from digestion by DNase I. Protected region I, from nucleotide +15 to nucleotide -10, spans the site of transcription initiation. Protected region II extends from nucleotide -70 to nucleotide -100 relative to initiation, falling within a 42-base-pair sequence which is homologous to the 60/81-base-pair repeated elements which occur outside of the promoter in the spacer. Protected region III is upstream of region II, from nucleotide -120 to nucleotide -140. All three regions correlate with sequences known from deletion studies to be important for promoter function. Deletion mutants which retain either region I or regions II and III together footprint normally. Deletion of region III, however, reduces but does not eliminate footprinting on region II, suggesting either that one protein binds to both regions or that the proteins which bind to these sites interact with each other.

摘要

从非洲爪蟾卵母细胞制备的提取物含有一种或多种蛋白质,这些蛋白质能特异性保护RNA聚合酶I启动子的三个离散区域不被DNase I消化。受保护区域I,从核苷酸+15到核苷酸-10,跨越转录起始位点。相对于起始位点,受保护区域II从核苷酸-70延伸到核苷酸-100,位于一个42个碱基对的序列内,该序列与间隔区中启动子外出现的60/81个碱基对的重复元件同源。受保护区域III在区域II的上游,从核苷酸-120到核苷酸-140。所有这三个区域都与缺失研究中已知的对启动子功能很重要的序列相关。保留区域I或区域II和III的缺失突变体正常产生足迹。然而,区域III的缺失会减少但不会消除区域II上的足迹,这表明要么一种蛋白质与这两个区域结合,要么与这些位点结合的蛋白质相互作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e240/366714/e1592c663037/molcellb00098-0051-a.jpg

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