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在不同生长条件下大肠杆菌稳定RNA操纵子的FIS依赖性反式激活。

FIS-dependent trans activation of stable RNA operons of Escherichia coli under various growth conditions.

作者信息

Nilsson L, Verbeek H, Vijgenboom E, van Drunen C, Vanet A, Bosch L

机构信息

Department of Biochemistry, Leiden University, The Netherlands.

出版信息

J Bacteriol. 1992 Feb;174(3):921-9. doi: 10.1128/jb.174.3.921-929.1992.

Abstract

In Escherichia coli transcription of the tRNA operon thrU (tufB) and the rRNA operon rrnB is trans-activated by the protein FIS. This protein, which stimulates the inversion of various viral DNA segments, binds specifically to a cis-acting sequence (designated UAS) upstream of the promoter of thrU (tufB) and the P1 promoter of the rrnB operon. There are indications that this type of regulation is representative for the regulation of more stable RNA operons. In the present investigation we have studied UAS-dependent transcription activation of the thrU (tufB) operon in the presence and absence of FIS during a normal bacterial growth cycle and after a nutritional shift-up. In early log phase the expression of the operon rises steeply in wild-type cells, whereafter it declines. Concomitantly, a peak of the cellular FIS concentration is observed. Cells in the stationary phase are depleted of FIS. The rather abrupt increase of transcription activation depends on the nutritional quality of the medium. It is not seen in minimal medium. After a shift from minimal to rich medium, a peak of transcription activation and of FIS concentration is measured. This peak gets higher as the medium gets more strongly enriched. We conclude that a correlation between changes of the UAS-dependent activation of the thrU (tufB) operon and changes of the cellular FIS concentration under a variety of experimental conditions exists. This correlation strongly suggests that the production of FIS responds to environmental signals, thereby trans-activating the operon. Cells unable to produce FIS (fis cells) also show an increase of operon transcription in the early log phase and after a nutritional shift-up, albeit less pronounced than that wild-type cells. Presumably it is controlled by the ribosome feedback regulatory system. cis activation of the operon by the upstream activator sequence is apparent in the absence of FIS. This activation is constant throughout the entire growth cycle and is independent of nutritional factors. The well-known growth rate-dependent control, displayed by exponentially growing cells studied under various nutritional conditions, is governed by two regulatory mechanisms: repression, presumably by ribosome feedback inhibition, and stimulation by trans activation. FIS allows very fast bacterial growth.

摘要

在大肠杆菌中,tRNA操纵子thrU(tufB)和rRNA操纵子rrnB的转录由蛋白质FIS反式激活。这种能刺激各种病毒DNA片段倒位的蛋白质,特异性结合在thrU(tufB)启动子上游的顺式作用序列(称为UAS)以及rrnB操纵子的P1启动子上。有迹象表明,这种调控类型代表了对更稳定RNA操纵子的调控。在本研究中,我们研究了在正常细菌生长周期以及营养上调后,存在和不存在FIS时thrU(tufB)操纵子的UAS依赖性转录激活。在对数早期,操纵子在野生型细胞中的表达急剧上升,随后下降。同时,观察到细胞FIS浓度出现峰值。稳定期的细胞中FIS耗尽。转录激活的相当突然的增加取决于培养基的营养质量。在基本培养基中未观察到这种情况。从基本培养基转变为丰富培养基后,测量到转录激活和FIS浓度的峰值。随着培养基富集程度越高,这个峰值越高。我们得出结论,在各种实验条件下,thrU(tufB)操纵子的UAS依赖性激活变化与细胞FIS浓度变化之间存在相关性。这种相关性强烈表明,FIS的产生对环境信号作出反应,从而反式激活操纵子。无法产生FIS的细胞(fis细胞)在对数早期和营养上调后也显示出操纵子转录增加,尽管不如野生型细胞明显。推测它受核糖体反馈调节系统控制。在没有FIS的情况下,上游激活序列对操纵子的顺式激活是明显的。这种激活在整个生长周期中是恒定的,并且与营养因素无关。在各种营养条件下研究的指数生长细胞所表现出的众所周知的生长速率依赖性控制,由两种调节机制控制:可能通过核糖体反馈抑制进行阻遏,以及通过反式激活进行刺激。FIS允许细菌非常快速地生长。

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