Young R A, Steitz J A
Proc Natl Acad Sci U S A. 1978 Aug;75(8):3593-7. doi: 10.1073/pnas.75.8.3593.
The nucleotide sequence of Escherichia coli DNA at both ends of the gene for 16S rRNA has been determined for two rRNA operons, rrnD and rrnX. The 400 nucleotides we have examined exhibit only one base change between rrnD and rrnX. Within the 160 nucleotides that precede mature 16S rRNA sequences are cleavage sites for several E. coli endonucleases, including RNase III. A 240-nucleotide segment encompassing the 16S 3' end contains another RNase III site and the point of presumed RNase P scission at the 5' end of tRNA1Ile, the first tRNA appearing in the 16-23S spacer region of rrnD and rrnX. Most importantly, the DNA sequences predict that regions flanking the 16S gene in the rRNA primary transcript extensively base pair to form a double-helical structure whose hairpin loop includes the entire mature 16S molecule; within this structure is a 26-base-pair stem containing the two sequences at which RNase III action generates the 5' and 3' ends of a previously characterized precursor to 16S rRNA. Although our proposed secondary structure for this RNase III site is superficially dissimilar to previously described cleavage sites in the T7 early mRNA precursor, certain common features may constitute signals for RNase III recognition. The suggestion that distant portions of an RNA molecule can form a secondary structure within which specific endonucleolytic cleavages occur may have mechanistic implications for the joining of noncontiguous portions of gene sequences evident in several eukaryotic mRNAs.
已确定了大肠杆菌rrnD和rrnX两个rRNA操纵子中16S rRNA基因两端的核苷酸序列。我们检测的400个核苷酸在rrnD和rrnX之间仅显示一个碱基变化。在成熟16S rRNA序列之前的160个核苷酸内,有几种大肠杆菌内切核酸酶的切割位点,包括RNase III。一个包含16S 3'末端的240个核苷酸片段含有另一个RNase III位点以及tRNA1Ile 5'末端假定的RNase P切割点,tRNA1Ile是rrnD和rrnX的16 - 23S间隔区中出现的第一个tRNA。最重要的是,DNA序列预测rRNA初级转录本中16S基因侧翼区域广泛碱基配对形成双螺旋结构,其发夹环包含整个成熟16S分子;在这个结构中有一个26个碱基对的茎,包含两个序列,在这两个序列处RNase III的作用产生了先前已鉴定的16S rRNA前体的5'和3'末端。虽然我们提出的这个RNase III位点的二级结构表面上与T7早期mRNA前体中先前描述的切割位点不同,但某些共同特征可能构成RNase III识别的信号。RNA分子的远距离部分可以形成二级结构,在其中发生特定的内切核酸酶切割,这一观点可能对几个真核mRNA中明显的基因序列非连续部分的连接具有机制上的启示。