卡尔斯伯酵母rRNA操纵子转录的体内和体外起始位点。

Klootwijk J, Verbeet M P, Veldman G M, de Regt V C, van Heerikhuizen H, Bogerd J, Planta R J

Nucleic Acids Res. 1984 Feb 10;12(3):1377-90. doi: 10.1093/nar/12.3.1377.

We have performed a detailed analysis of the transcription initiation of the rRNA operon in the yeast Saccharomyces carlsbergensis. Electron microscopic analysis of R-looped pre-rRNA molecules together with a very sensitive S1-nuclease mapping showed the use of only a single transcription start at about 700 bp upstream of the 17S rRNA gene and not of the minor start sites proposed for the very closely related species S. cerevisiae by others [Bayev et al. (5), Swanson and Holland (6)]. The sequence of 730 bp of the initiating region is presented. In vitro transcription in concentrated lysates of yeast spheroplasts in the presence of (gamma-SH)ATP or (gamma-SH)GTP, followed by purification of the in vitro initiated RNA via Hg-agarose, revealed that on the endogenous template exactly the same site is used for transcription initiation as in vivo.

我们已经对卡尔斯伯酵母中rRNA操纵子的转录起始进行了详细分析。对R环前体rRNA分子进行电子显微镜分析，并结合非常灵敏的S1核酸酶图谱分析，结果表明，转录起始仅使用17S rRNA基因上游约700 bp处的一个位点，而不是其他研究人员针对密切相关的酿酒酵母提出的次要起始位点[Bayev等人(5)，Swanson和Holland(6)]。本文给出了起始区域730 bp的序列。在存在(γ-SH)ATP或(γ-SH)GTP的情况下，在酵母原生质体浓缩裂解物中进行体外转录，然后通过汞琼脂糖纯化体外起始的RNA，结果显示，在内源模板上，转录起始位点与体内完全相同。