Institute of Biochemistry and Biophysics, Polish Academy of Sciences, 02-106 Warsaw, Poland.
Int J Mol Sci. 2021 May 11;22(10):5066. doi: 10.3390/ijms22105066.
encodes a large set of transcriptional regulators (TRs) that modulate and manage cellular metabolism to survive in variable environmental conditions including that of the human body. The AraC family regulators are an abundant group of TRs in bacteria, mostly acting as gene expression activators, controlling diverse cellular functions (e.g., carbon metabolism, stress response, and virulence). The PA3027 protein from has been classified in silico as a putative AraC-type TR. Transcriptional profiling of PAO1161 overexpressing revealed a spectacular increase in the mRNA levels of - (divergent to ), , and genes encoding proteins potentially involved in glycerolipid metabolism. Concomitantly, chromatin immunoprecipitation-sequencing (ChIP-seq) analysis revealed that at least 22 regions are bound by PA3027 in the PAO1161 genome. These encompass promoter regions of , , and , showing the major increase in expression in response to PA3027 excess. In Vitro DNA binding assay confirmed interactions of PA3027 with these regions. Furthermore, promoter-reporter assays in a heterologous host showed the PA3027-dependent activation of the promoter of the - operon. Two motifs representing the preferred binding sites for PA3027, one localized upstream and one overlapping with the -35 promoter sequence, were identified in p and our data indicate that both motifs are required for full activation of this promoter by PA3027. Overall, the presented data show that PA3027 acts as a transcriptional regulator in , activating genes likely engaged in glycerolipid metabolism. The GliR name, from a glycerolipid metabolism regulator, is proposed for PA3027 of .
编码了一大组转录调节剂 (TRs),这些调节剂可以调节和管理细胞代谢,以在包括人体在内的各种环境条件下生存。AraC 家族调节剂是细菌中大量的 TRs 群体,主要作为基因表达激活剂,控制多种细胞功能(例如,碳代谢、应激反应和毒力)。来自 的 PA3027 蛋白已在计算机中被分类为假定的 AraC 型 TR。 过表达的 的转录谱显示,编码潜在参与甘油脂质代谢的蛋白质的 、 、和 基因的 mRNA 水平显着增加。同时,染色质免疫沉淀测序 (ChIP-seq) 分析显示,PA3027 在 PAO1161 基因组中至少结合了 22 个区域。这些区域包含 、 和 的启动子区域,这些区域的表达在 PA3027 过量时主要增加。体外 DNA 结合测定证实了 PA3027 与这些区域的相互作用。此外,在异源宿主中的启动子报告基因测定显示,PA3027 依赖于 - 操纵子启动子的激活。在 p 中鉴定了两个代表 PA3027 优先结合位点的基序,一个位于上游,一个与 -35 启动子序列重叠,并且我们的数据表明这两个基序对于 PA3027 对该启动子的完全激活都是必需的。总体而言,所呈现的数据表明,PA3027 在 中作为转录调节剂发挥作用,激活可能参与甘油脂质代谢的基因。拟议将 GliR 名称赋予 中的 PA3027,用于甘油脂质代谢调节剂。