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[大肠杆菌hns基因座内的反义转录]

[Antisense transcription within the hns locus of Escherichia coli].

作者信息

Tututkina M N, Shavkunov K S, Masulis I S, Ozolin' O N

出版信息

Mol Biol (Mosk). 2010 May-Jun;44(3):497-506.

Abstract

Scanning the entire genome of E. coli by means of pattern-recognition software PlatProm spotted out more than a thousand of potential start points for antisense transcription. Taking into account possible role of antisense RNAs in the cell regulatory networks, our top-priority interest was focused on the promoter-like sites found within genes of transcription regulators. One of them (hns) encodes a major nucleoid protein affecting expression pattern of many genomic loci. Several potential start points for antisense transcription were found within its coding sequence. Gel-retardation assays, potassium permanganate and DNAse I foot-printings confirmed the ability of the intragenic promoter located approximately 280 bp downstream of ATG to bind RNA polymerase. Primer extension revealed the cDNA of the expected size while Northern blot hybridization assumes the presence of aRNA among cellular RNAs. Relative abundance of antisense RNA and hns-mRNA in vivo exhibited dependence on growth conditions thus assuming existence of regulatory pathways keeping cellular concentration of these two transcripts at the optimal level.

摘要

通过模式识别软件PlatProm扫描大肠杆菌的全基因组,发现了一千多个潜在的反义转录起始点。考虑到反义RNA在细胞调控网络中的可能作用,我们的首要兴趣集中在转录调节因子基因内发现的启动子样位点。其中一个(hns)编码一种主要的类核蛋白,影响许多基因组位点的表达模式。在其编码序列内发现了几个潜在的反义转录起始点。凝胶阻滞分析、高锰酸钾和DNA酶I足迹分析证实,位于ATG下游约280 bp处的基因内启动子能够结合RNA聚合酶。引物延伸揭示了预期大小的cDNA,而Northern印迹杂交表明细胞RNA中存在反义RNA。体内反义RNA和hns-mRNA的相对丰度表现出对生长条件的依赖性,因此推测存在调控途径将这两种转录本的细胞浓度维持在最佳水平。

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