Engberg J, Din N, Saiga H, Higashinakagawa T
Nucleic Acids Res. 1984 Jan 25;12(2):959-72. doi: 10.1093/nar/12.2.959.
The 5'-terminus of 35S pre-rRNA and mature 17S rRNA of Tetrahymena thermophila was mapped on cloned rDNA fragments by S1 nuclease protection experiments. A single site for transcription initiation was observed when pre-rRNA prepared by three different methods was used as RNA probe. These mapping results were unambiguously confirmed by sequencing the 5'-terminal region of in vitro capped 35S pre-rRNA. DNA sequence analysis of about 520 nucleotides upstream of the transcription initiation site revealed several distinct sets of highly conserved repeat sequences. In addition, the 840 nucleotides downstream of the transcription initiation site (+ 1) was determined and shown to include the 5'-terminus of the 17S rRNA coding region at position + 647. A region surrounding the position + 195 contains an inverted repeat sequence which could be the structural basis for the recently described premature transcription termination event in this organism (Kister et al. (1983) Nucl. Acids Res. 11, 3487-3502).
嗜热四膜虫35S前体rRNA和成熟17S rRNA的5′端通过S1核酸酶保护实验定位在克隆的rDNA片段上。当使用三种不同方法制备的前体rRNA作为RNA探针时,观察到一个单一的转录起始位点。通过对体外加帽的35S前体rRNA的5′端区域进行测序,明确证实了这些定位结果。对转录起始位点上游约520个核苷酸的DNA序列分析揭示了几组不同的高度保守重复序列。此外,确定了转录起始位点(+1)下游的840个核苷酸,并显示其在+647位置包含17S rRNA编码区的5′端。+195位置周围的区域包含一个反向重复序列,这可能是该生物体中最近描述的过早转录终止事件的结构基础(基斯特等人(1983年)《核酸研究》11,3487 - 3502)。