卡氏棘阿米巴核糖体RNA转录单位的起始位点。

The start site of the Acanthamoeba castellanii ribosomal RNA transcription unit.

作者信息

Perna P J, Harris G H, Iida C T, Kownin P, Bugren S, Paule M R

机构信息

Department of Biochemistry, Colorado State University, Fort Collins 80523.

出版信息

Gene Expr. 1992;2(1):71-8.

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6057357/

Abstract

The 39S ribosomal RNA (rRNA) precursor has been isolated from Acanthamoeba castellanii. In vitro capping of the isolated RNA verified that it is the primary transcript and identified the 5' nucleotide as pppA. The position of the 5' coding nucleotide on the rRNA repeat unit sequence was identified using Northern blot, R-loop, and S1 nuclease mapping techniques. Dinucleotide priming of an in vitro transcription system stalled because of low initiating nucleotide concentration revealed that ApA maximally stimulates initiation of transcription. All of these results show that the underlined A in the sequence 5'-TATATATAAAGGGAC (RNA-like strand) coincides with the 5' nucleotide of the primary transcript. This identification is compatible with in vitro transcription experiments mapping the promoter for this transcription unit. The initiation sequences of rRNA genes from 14 species are compared, and a weak consensus for the initiator derived: [Formula; see text].

摘要

已从卡氏棘阿米巴中分离出39S核糖体RNA（rRNA）前体。对分离出的RNA进行体外加帽验证，证实其为初级转录本，并确定5'核苷酸为pppA。使用Northern印迹、R环和S1核酸酶作图技术确定了rRNA重复单元序列上5'编码核苷酸的位置。体外转录系统由于起始核苷酸浓度低而导致二核苷酸引发停滞，结果表明ApA最大程度地刺激转录起始。所有这些结果表明，序列5'-TATATATAAAGGGAC（RNA样链）中带下划线的A与初级转录本的5'核苷酸一致。这一鉴定结果与绘制该转录单元启动子的体外转录实验结果相符。比较了14个物种的rRNA基因的起始序列，并得出了起始子的弱共有序列：[公式；见正文] 。

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