School of Biological Sciences, Department of Molecular Biology, University of California-San Diego, La Jolla, CA 92093, USA.
Int J Mol Sci. 2022 Sep 7;23(18):10343. doi: 10.3390/ijms231810343.
Using reporter gene () transcriptional fusions we examined the transcriptional dependencies of the promoter (P) and the entire operon regulatory region (P-) on eight transcription factors as well as the inducer, salicin, and an IS5 insertion upstream of P. Crp-cAMP is the primary activator of both P and the operon, while H-NS is a strong dominant operon repressor but only a weak repressor of P. H-NS may exert its repressive effect by looping the DNA at two binding sites. StpA is a relatively weak repressor in the absence of H-NS, while Fis also has a weak repressive effect. Salicin has no effect on P activity but causes a 30-fold induction of operon expression. Induction depends on the activity of the BglF transporter/kinase. IS5 insertion has only a moderate effect on P but causes a much greater activation of the operon expression by preventing the full repressive effects of H-NS and StpA. While several other transcription factors (BglJ, RcsB, and LeuO) have been reported to influence operon transcription when overexpressed, they had little or no effect when present at wild type levels. These results indicate the important transcriptional regulatory mechanisms operative on the operon in .
利用报告基因()转录融合,我们研究了启动子(P)和整个操纵子调控区(P-)对八个转录因子以及诱导物水杨苷和 P 上游的 IS5 插入的转录依赖性。Crp-cAMP 是 P 和 操纵子的主要激活剂,而 H-NS 是一个强大的操纵子抑制剂,但对 P 的抑制作用较弱。H-NS 可能通过在两个结合位点处使 DNA 环化来发挥其抑制作用。在没有 H-NS 的情况下,StpA 是一个相对较弱的抑制剂,而 Fis 也有较弱的抑制作用。水杨苷对 P 活性没有影响,但会导致 操纵子表达的 30 倍诱导。诱导依赖于 BglF 转运体/激酶的活性。IS5 插入对 P 的影响不大,但通过防止 H-NS 和 StpA 的完全抑制作用,大大激活了 操纵子的表达。虽然已经报道了其他几个转录因子(BglJ、RcsB 和 LeuO)在过表达时会影响 操纵子转录,但在野生型水平时它们几乎没有或没有影响。这些结果表明了 在 中起作用的重要转录调控机制。