Rachwalski Kenneth, Madden Sean J, Ritchie Nicole, French Shawn, Bhando Timsy, Girgis-Gabardo Adele, Tu Megan, Gordzevich Rodion, Ives Rowan, Guo Amelia B Y, Johnson Jarrod W, Xu Yiming, Kapadia Sharookh B, Magolan Jakob, Brown Eric D
Institute of Infectious Disease Research, McMaster University, Hamilton, ON L8S 4L8, Canada.
Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON L8S 4L8, Canada.
iScience. 2024 Sep 5;27(10):110894. doi: 10.1016/j.isci.2024.110894. eCollection 2024 Oct 18.
The increasing prevalence of antibiotic resistance demands the discovery of antibacterial chemical scaffolds with unique mechanisms of action. Phenotypic screening approaches, such as the use of reporters for bacterial cell stress, offer promise to identify compounds while providing strong hypotheses for follow-on mechanism of action studies. From a collection of ∼1,800 GFP transcriptional reporter strains, we identified a reporter that is highly induced by cell envelope stress-pProm -GFP. After characterizing pProm -GFP induction, we assessed a collection of bioactive small molecules for reporter induction, identifying 24 compounds of interest. Spontaneous suppressors to one compound in particular, MAC-0452936, mapped to the gene encoding the essential prolipoprotein diacylglyceryl transferase, . Lgt inhibition by MAC-0452936 inhibition was confirmed through genetic, phenotypic, and biochemical approaches. The oxime ester, MAC-0452936, represents a useful small molecule inhibitor of Lgt and highlights the potential of using pProm -GFP as a phenotypic screening tool.
抗生素耐药性的日益普遍，需要发现具有独特作用机制的抗菌化学骨架。表型筛选方法，如使用细菌细胞应激报告基因，有望识别化合物，同时为后续的作用机制研究提供有力假设。从大约1800个绿色荧光蛋白（GFP）转录报告菌株库中，我们鉴定出一个受细胞包膜应激高度诱导的报告基因——pProm -GFP。在对pProm -GFP的诱导进行表征后，我们评估了一系列生物活性小分子对报告基因的诱导作用，鉴定出24种感兴趣的化合物。特别是对一种化合物MAC-0452936的自发抑制子，定位到编码必需的前脂蛋白二酰甘油转移酶Lgt的基因。通过遗传学、表型和生化方法证实了MAC-0452936对Lgt的抑制作用。肟酯MAC-0452936是一种有用的Lgt小分子抑制剂，突出了使用pProm -GFP作为表型筛选工具的潜力。
