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ArgR调节蛋白,在铜绿假单胞菌中arcD启动子转录激活过程中作为厌氧调节因子ANR的辅助因子。

The ArgR regulatory protein, a helper to the anaerobic regulator ANR during transcriptional activation of the arcD promoter in Pseudomonas aeruginosa.

作者信息

Lu C D, Winteler H, Abdelal A, Haas D

机构信息

Department of Biology, Georgia State University, Atlanta, Georgia 30303, USA.

出版信息

J Bacteriol. 1999 Apr;181(8):2459-64. doi: 10.1128/JB.181.8.2459-2464.1999.

Abstract

Pseudomonas aeruginosa, when deprived of oxygen, generates ATP from arginine catabolism by enzymes of the arginine deiminase pathway, encoded by the arcDABC operon. Under conditions of low oxygen tension, the transcriptional activator ANR binds to a site centered 41.5 bp upstream of the arcD transcriptional start. ANR-mediated anaerobic induction was enhanced two- to threefold by extracellular arginine. This arginine effect depended, in trans, on the transcriptional regulator ArgR and, in cis, on an ArgR binding site centered at -73.5 bp in the arcD promoter. Binding of purified ArgR protein to this site was demonstrated by electrophoretic mobility shift assays and DNase I footprinting. This ArgR recognition site contained a sequence, 5'-TGACGC-3', which deviated in only 1 base from the common sequence motif 5'-TGTCGC-3' found in other ArgR binding sites of P. aeruginosa. Furthermore, an alignment of all known ArgR binding sites confirmed that they consist of two directly repeated half-sites. In the absence of ANR, arginine did not induce the arc operon, suggesting that ArgR alone does not activate the arcD promoter. According to a model proposed, ArgR makes physical contact with ANR and thereby facilitates initiation of arc transcription.

摘要

铜绿假单胞菌在缺氧时,通过精氨酸脱亚胺酶途径的酶从精氨酸分解代谢中产生ATP,该途径由arcDABC操纵子编码。在低氧张力条件下,转录激活因子ANR结合到arcD转录起始上游41.5 bp处的一个位点。细胞外精氨酸使ANR介导的厌氧诱导增强两到三倍。这种精氨酸效应反式依赖于转录调节因子ArgR,顺式依赖于arcD启动子中位于 -73.5 bp处的一个ArgR结合位点。通过电泳迁移率变动分析和DNase I足迹实验证明了纯化的ArgR蛋白与该位点的结合。这个ArgR识别位点包含一个序列5'-TGACGC-3',与铜绿假单胞菌其他ArgR结合位点中发现的共有序列基序5'-TGTCGC-3'仅相差1个碱基。此外,所有已知的ArgR结合位点的比对证实它们由两个直接重复的半位点组成。在没有ANR的情况下,精氨酸不会诱导arc操纵子,这表明单独的ArgR不会激活arcD启动子。根据提出的一个模型,ArgR与ANR进行物理接触,从而促进arc转录的起始。

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