Chattopadhyay S, Wu Y, Datta P
Department of Biological Chemistry, The University of Michigan, Ann Arbor 48109-0606, USA.
J Bacteriol. 1997 Aug;179(15):4868-73. doi: 10.1128/jb.179.15.4868-4873.1997.
Anaerobic expression of the tdcABC operon in Escherichia coli, as measured by LacZ activity from single-copy tdc-lacZ transcriptional and translational fusions, is greatly reduced in strains lacking two global transcriptional regulators, Fnr and ArcA. The nucleotide sequence of the tdc promoter around -145 shows significant similarity with the consensus Fnr-binding site; however, extensive base substitutions within this region had no effect on Fnr regulation of the tdc genes. A genetic analysis revealed that the effect of Fnr on tdc is not mediated via ArcA. Furthermore, addition of cyclic AMP to the anaerobic incubation medium completely restored tdc expression in fnr and arcA mutants as well as in strains harboring mutations in the Fnr- and ArcA-dependent pfl gene and the Fnr-regulated glpA and frd genes. These results, taken together with the earlier finding that tdc expression is subject to catabolite repression by intermediary metabolites, strongly suggest that the negative regulatory effects of mutations in the fnr and arcA genes are mediated physiologically due to accumulation of a metabolite(s) which prevents tdc transcription in vivo.
通过单拷贝tdc - lacZ转录和翻译融合体的LacZ活性测定,在大肠杆菌中tdcABC操纵子的厌氧表达在缺乏两种全局转录调节因子Fnr和ArcA的菌株中大大降低。tdc启动子在-145附近的核苷酸序列与Fnr结合位点的共有序列具有显著相似性;然而,该区域内的广泛碱基替换对tdc基因的Fnr调节没有影响。遗传分析表明,Fnr对tdc的作用不是通过ArcA介导的。此外,向厌氧培养培养基中添加环磷酸腺苷完全恢复了fnr和arcA突变体以及在Fnr和ArcA依赖性pfl基因以及Fnr调节的glpA和frd基因中携带突变的菌株中的tdc表达。这些结果与早期发现tdc表达受中间代谢物的分解代谢物阻遏的结果一起,强烈表明fnr和arcA基因中的突变的负调节作用在生理上是由于一种代谢物的积累介导的,这种代谢物在体内阻止tdc转录。