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对噬菌体λ上第三个依赖于cII且与溶源发育相关的协同激活启动子的表征。

Characterization of a third, cII-dependent, coordinately activated promoter on phage lambda involved in lysogenic development.

作者信息

Ho Y S, Rosenberg M

出版信息

J Biol Chem. 1985 Sep 25;260(21):11838-44.

PMID:2931430
Abstract

Lysogenic development by bacteriophage lambda is known to require the coordinate expression of two phage operons. Coordinate control is achieved by a positive regulatory mechanism which activates transcription from the promoters of these operons, PRE and PI. The positive effector is the phage regulatory protein cII. We now identify and characterize a third cII-dependent transcription unit on phage lambda, which is positioned in the middle of the Q regulatory gene and has an anti-Q orientation. We demonstrate the cII-dependent function of this promoter and precisely map its 5' transcription start-site both in vitro and in vivo. Most importantly, we show that cII binding and transcription activation at PaQ occur at essentially the same cII levels as those required for PRE and PI activation, and that all three promoters respond to cII at the same time following phage infection. In addition, DNase protection studies demonstrate that cII selectively interacts with the same TTGCN6TTGC DNA sequence repeat in the -35 region of PaQ which cII interacts with at both PRE and PI. We find that cII also binds other TTGCN6TTGC repeat sequences on lambda but binding at these sites does not lead to productive transcription. We conclude that PaQ functions in concert with PRE and PI to regulate the lysogenic growth response of phage lambda. We presume that the PaQ directed anti-sense Q RNA transcript functions to down-regulate the expression of the Q gene, which is needed for the expression of all phage late genes during lytic growth.

摘要

已知噬菌体λ的溶原性发育需要两个噬菌体操纵子的协同表达。协同控制是通过一种正调控机制实现的,该机制激活这些操纵子的启动子PRE和PI的转录。正效应物是噬菌体调控蛋白cII。我们现在鉴定并表征了噬菌体λ上的第三个cII依赖性转录单元,它位于Q调控基因的中间,并且具有反Q方向。我们证明了该启动子的cII依赖性功能,并在体外和体内精确绘制了其5'转录起始位点。最重要的是,我们表明在PaQ处的cII结合和转录激活发生时的cII水平与PRE和PI激活所需的水平基本相同,并且在噬菌体感染后所有三个启动子同时对cII作出反应。此外,DNA酶保护研究表明,cII在PaQ的 -35区域与相同的TTGCN6TTGC DNA序列重复选择性相互作用,cII在PRE和PI处也与该序列相互作用。我们发现cII也结合λ上的其他TTGCN6TTGC重复序列,但在这些位点的结合不会导致有效的转录。我们得出结论,PaQ与PRE和PI协同作用以调节噬菌体λ的溶原性生长反应。我们推测由PaQ指导的反义Q RNA转录本的功能是下调Q基因的表达,而Q基因的表达是裂解生长期间所有噬菌体晚期基因表达所必需的。

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