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Negative Transcriptional Regulation of the ilv-leu Operon for Biosynthesis of Branched-Chain Amino Acids through the Bacillus subtilis Global Regulator TnrA

机译:通过枯草芽孢杆菌全局调节剂TnrA的支链氨基酸生物合成的ilv-leu操纵子的负转录调控

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摘要

The Bacillus subtilis ilv-leu operon is involved in the synthesis of branched-chain amino acids (valine, isoleucine, and leucine). The two- to threefold repression of expression of the ilv-leu operon during logarithmic-phase growth under nitrogen-limited conditions, which was originally detected by a DNA microarray analysis to compare the transcriptomes from the wild-type and tnrA mutant strains, was confirmed by lacZ fusion and Northern experiments. A genome-wide TnrA box search revealed a candidate box approximately 200 bp upstream of the transcription initiation base of the ilv-leu operon, the TnrA binding to which was verified by gel retardation and DNase I footprinting analyses. Deletion and base substitution of the TnrA box sequence affected the ilv-leu promoter activity in vivo, implying that TnrA bound to the box might be able to inhibit the promoter activity, possibly through DNA bending. The negative control of the expression of the ilv-leu operon by TnrA, which is considered to represent rather fine-tuning (two- to threefold), is a novel regulatory link between nitrogen and amino acid metabolism.
机译:枯草芽孢杆菌ilv-leu操纵子参与分支链氨基酸(缬氨酸,异亮氨酸和亮氨酸)的合成。证实了在氮限制条件下对数生长期中ilv-leu操纵子表达的2至3倍抑制,该抑制最初是通过DNA芯片分析检测以比较野生型和tnrA突变菌株的转录组而检测到的。通过lacZ融合和Northern实验。全基因组的TnrA框搜索显示了ilv-leu操纵子转录起始碱基上游约200 bp的候选框,通过凝胶阻滞和DNase I足迹分析验证了与TnrA的结合。 TnrA盒序列的缺失和碱基取代影响了体内ilv-leu启动子的活性,这意味着与该盒结合的TnrA可能能够抑制启动子的活性,可能是通过DNA弯曲。 TnrA对ilv-leu操纵子表达的负控制被认为代表了微调(2到3倍),是氮与氨基酸代谢之间的新型调控环节。

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