In the search for much-needed new antibacterial chemical matter, a myriad of compounds have been reported in academic and pharmaceutical screening endeavors. Only a small fraction of these, however, are characterized with respect to mechanism of action (MOA). Here, we describe a pipeline that categorizes transcriptional responses to antibiotics and provides hypotheses for MOA. 3D-printed imaging hardware PFIboxes) profiles responses of promoter-GFP fusions to more than 100 antibiotics. Notably, metergoline, a semi-synthetic ergot alkaloid, mimics a DNA replication inhibitor. supercoiling assays confirm this prediction, and a potent analog thereof (MLEB-1934) inhibits growth at 0.25 μg/mL and is highly active against quinolone-resistant strains of methicillin-resistant . Spontaneous suppressor mutants map to a seldom explored allosteric binding pocket, suggesting a mechanism distinct from DNA gyrase inhibitors used in the clinic. In all, the work highlights the potential of this platform to rapidly assess MOA of new antibacterial compounds.

中文翻译：

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基于细菌转录反应预测作用机制的平台鉴定出一种不寻常的 DNA 旋转酶抑制剂


在寻找急需的新型抗菌化学物质的过程中，学术和药物筛选工作中报道了无数的化合物。然而，其中只有一小部分具有作用机制 (MOA) 的特征。在这里，我们描述了一个对抗生素转录反应进行分类并为 MOA 提供假设的管道。 3D 打印成像硬件 PFIboxes）描绘了启动子-GFP 融合体对 100 多种抗生素的反应。值得注意的是，麦角林是一种半合成麦角生物碱，可模仿 DNA 复制抑制剂。超螺旋测定证实了这一预测，其有效类似物 (MLEB-1934) 在 0.25 μg/mL 下抑制生长，并且对耐喹诺酮菌株和耐甲氧西林菌株具有高度活性。自发抑制突变体映射到一个很少探索的变构结合口袋，这表明其机制不同于临床中使用的 DNA 旋转酶抑制剂。总之，这项工作凸显了该平台快速评估新型抗菌化合物 MOA 的潜力。