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大肠杆菌中tgt/sec操纵子的启动子之前有一个上游激活序列，该序列包含一个高亲和力的FIS结合位点。

The promoter of the tgt/sec operon in Escherichia coli is preceded by an upstream activation sequence that contains a high affinity FIS binding site.

作者信息

Slany R K, Kersten H

机构信息

Institut für Biochemie, Universität Erlangen-Nürnberg, Germany.

出版信息

Nucleic Acids Res. 1992 Aug 25;20(16):4193-8. doi: 10.1093/nar/20.16.4193.

DOI:10.1093/nar/20.16.4193
PMID:1508713
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC334125/
Abstract

The tgt/sec operon in E. coli consists of five genes: queA, tgt, ORF12, secD, and secF. QueA and Tgt participate in the biosynthesis of the hypermodified t-RNA nucleoside Queuosine, whereas SecD and SecF are involved in protein secretion. Examination of the promoter region of the operon showed structural similarity to promoter regions of the rrn-operons. An upstream activation sequence (UAS) containing a potential binding site for the factor of inversion stimulation (FIS) was found. Gel retardation assays and DNaseI footprinting indicated, that FIS binds specifically and with high affinity to a site centred at position -58. Binding of FIS caused bending of the DNA, as deduced from circular permutation analysis. Various 5' deletion mutants of the promoter region were constructed and fused to a lacZ reporter gene to determine the influence of the UAS element on the promoter strength. An approximately two-fold activation of the promoter by the UAS element was observed.

摘要

大肠杆菌中的tgt/sec操纵子由五个基因组成：queA、tgt、ORF12、secD和secF。QueA和Tgt参与超修饰tRNA核苷queuosine的生物合成，而SecD和SecF则参与蛋白质分泌。对该操纵子启动子区域的检查显示，其与rrn操纵子的启动子区域具有结构相似性。发现了一个上游激活序列（UAS），其中包含一个潜在的倒位刺激因子（FIS）结合位点。凝胶阻滞分析和DNaseI足迹分析表明，FIS特异性且高亲和力地结合到以-58位为中心的位点。从环形置换分析推断，FIS的结合导致了DNA的弯曲。构建了启动子区域的各种5'缺失突变体，并将其与lacZ报告基因融合，以确定UAS元件对启动子强度的影响。观察到UAS元件对启动子有大约两倍的激活作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/824d/334125/e36c603c05f6/nar00227-0081-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/824d/334125/7a952bc45c9f/nar00227-0079-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/824d/334125/8c298f308265/nar00227-0079-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/824d/334125/d2c39e9a779c/nar00227-0080-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/824d/334125/e36c603c05f6/nar00227-0081-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/824d/334125/7a952bc45c9f/nar00227-0079-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/824d/334125/8c298f308265/nar00227-0079-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/824d/334125/d2c39e9a779c/nar00227-0080-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/824d/334125/e36c603c05f6/nar00227-0081-a.jpg

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