Wu A M, Platt T
Proc Natl Acad Sci U S A. 1978 Nov;75(11):5442-6. doi: 10.1073/pnas.75.11.5442.
We have determined the RNA and DNA sequences in the region specifying termination of transcription at the end of the tryptophan (trp) operon of Escherichia coli. A 3'-terminal mRNA fragment of about 150 nucleotides yielded oligonucleotide products that could be assigned to the end of trpA (the last structural gene in the operon) by correlation with the amino acid sequence of the protein product. Analysis of the DNA corresponding to this region served to align the few noncoding RNA oligonucleotide sequences and demonstrated that termination of trp transcription occurs in vivo at a site 36 nucleotides after trpA, with greater than 95% efficiency. In two different strains partially defective in the transcription termination factor rho, the purified transcript is much longer and more complex, suggesting that a significant amount of read-through occurs in these strains. This is consistent with evidence [Guarente, L. P., Mitchell, D. H. & Beckwith, J. (1977) J. Mol. Biol. 112, 423-436] that efficient termination in vivo at the end of the trp operon is a rho-dependent event. The trp terminator (trp t) shares several features with other known sites of transcription termination, including (i) a 3'-terminal RNA sequence of several uridine residues, C-A-U-U-U-U(OH), (ii) a G.C-rich region in the DNA immediately preceding the site of termination, followed by an A.T-rich region, and (iii) a region of dyad symmetry in the DNA which, in the transcript, is capable of forming a stable hairpin containing seven G.C base pairs and one A.U base pair in its stem.
我们已经确定了大肠杆菌色氨酸(trp)操纵子末端转录终止区域的RNA和DNA序列。一个约150个核苷酸的3'末端mRNA片段产生的寡核苷酸产物,通过与蛋白质产物的氨基酸序列相关联,可确定其对应于trpA(操纵子中的最后一个结构基因)的末端。对该区域相应DNA的分析用于排列少数非编码RNA寡核苷酸序列,并证明trp转录在体内于trpA下游36个核苷酸处的位点终止,终止效率大于95%。在转录终止因子rho部分缺陷的两种不同菌株中,纯化的转录本更长且更复杂,这表明在这些菌株中发生了大量的通读现象。这与[Guarente, L. P., Mitchell, D. H. & Beckwith, J. (1977) J. Mol. Biol. 112, 423 - 436]的证据一致,即trp操纵子末端在体内的有效终止是一个依赖rho的事件。trp终止子(trp t)与其他已知的转录终止位点有几个共同特征,包括:(i)3'末端有几个尿苷残基的RNA序列,C - A - U - U - U - U(OH);(ii)在终止位点之前紧邻的DNA中有一个富含G.C的区域,随后是一个富含A.T的区域;(iii)DNA中的一个二元对称区域,在转录本中能够形成一个稳定的发夹结构,其茎部含有七个G.C碱基对和一个A.U碱基对。