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首页> 外文期刊>Molecular and Cellular Biology >Protein-DNA cross-linking reveals dramatic variation in RNA polymerase II density on different histone repeats of Drosophila melanogaster.
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Protein-DNA cross-linking reveals dramatic variation in RNA polymerase II density on different histone repeats of Drosophila melanogaster.

机译：蛋白质-DNA交联显示RNA聚合酶II密度对不同组蛋白重复的RNA聚合酶II密度的显着变化。

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摘要

In Drosophila melanogaster the five histone genes are within a 5-kilobase region which is repeated 100 times at a single chromosomal site. These 5-kilobase repeats are of two distinct classes, short and long, that differ by approximately 200 base pairs of DNA in the spacer region between the H1 and H3 genes. Since the mRNA-homologous regions of the repeats are highly conserved, one cannot examine differential expression of the repeats by classical hybridization methods. In this study, we assessed their transcriptional activity by measuring in vivo the relative amounts of RNA polymerase II that were cross-linked by UV irradiation to the two different histone repeats. The RNA polymerase II density on the long repeat in Schneider line 2 cells was strikingly lower (10-fold) than the density on the short repeat. The magnitude of this difference cannot be accounted for by reduced transcription of only one or two genes of the repeat. The density of topoisomerase I, an indicator of transcriptional activity, was also much higher on the short repeat than on the long repeat of line 2 cells. In contrast, the RNA polymerase II density was slightly higher on the long repeat than on the short repeat in a second cell line, KcH. The major difference between active (KcH) and inactive (S2) long repeats resides in the H1-H3 nontranscribed spacer. This portion of the spacer may contain a component necessary for expression that can act over a moderate distance and affect multiple genes of the repeat.
机译：在果蝇黑色素体中，五个组蛋白基因在5千碱基区域内，在单个染色体位点重复100次。这些5千碱基重复具有两个不同的类，短期和长，在H1和H3基因之间的间隔区中的间隔区域中的大约200碱基对DNA不同。由于重复的mRNA - 同源区域是高度保守的，因此通过经典杂交方法无法检查重复的差异表达。在这项研究中，我们通过体内测量通过UV辐射与两个不同的组蛋白重复交联的RNA聚合酶II的相对量来评估转录活性。在施耐德管线2细胞中长重复的RNA聚合酶II密度比短重复的密度小得多（10倍）。不能通过减少重复的一个或两个基因的转录来计算这种差异的幅度。拓扑异构酶I，转录活性的指示剂的密度在短的重复时比长重复的线2细胞在短的重复上高得多。相反，RNA聚合酶II密度在长重复比第二细胞系Kch中的短重复略高。活性（KCH）和无效（S2）长重复之间的主要区别在于H1-H3非段落的间隔物。间隔物的该部分可含有表达所需的组分，其表达可以在中等距离上作用并影响重复的多个基因。

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