Andersen J T, Poulsen P, Jensen K F
Institute of Biological Chemistry B, University of Copenhagen, Denmark.
Eur J Biochem. 1992 Jun 1;206(2):381-90. doi: 10.1111/j.1432-1033.1992.tb16938.x.
We have substituted on a plasmid the native promoter of the Escherichia coli rph-pyrE operon with an inducible transcription-initiation signal. The plasmid was used to study the mRNA chains derived from the operon at different intracellular concentrations of UTP and as a function of time following induction of transcription. The results showed that dicistronic rph-pyrE mRNA was formed when the UTP pool was low, and that a monocistronic rph mRNa was the major transcription product in high-UTP pools, thus supporting an UTP-controlled attenuation mechanism for regulation of pyrE gene expression. However, the dicistronic rph-pyrE transcript was rapidly processed into two monocistronic mRNA units, and a cleavage site was mapped near the attenuator in the intercistronic region, close to the site where transcription was terminated in high-UTP pools. Furthermore, the major 3' end of the pyrE mRNA was mapped near a palindromic structure of similarity to the family of repetitive extragenic palindromic sequences, 35 nucleotide residues after stop codon of the pryE gene.
我们已在质粒上用可诱导转录起始信号替换了大肠杆菌rph-pyrE操纵子的天然启动子。该质粒用于研究在不同细胞内浓度的UTP条件下以及转录诱导后随时间变化源自该操纵子的mRNA链。结果表明,当UTP库较低时会形成双顺反子rph-pyrE mRNA,而单顺反子rph mRNA是高UTP库中的主要转录产物,从而支持了一种UTP控制的衰减机制来调节pyrE基因表达。然而,双顺反子rph-pyrE转录本会迅速加工成两个单顺反子mRNA单元,并且在顺反子间区域的衰减子附近定位了一个切割位点,该位点靠近高UTP库中转录终止的位点。此外,pyrE mRNA的主要3'端定位于与重复基因外回文序列家族相似的回文结构附近,在pyrE基因终止密码子后35个核苷酸残基处。