High-throughput identification of the sensitivities of an Escherichia coli ΔrecA mutant strain to various chemical compounds
High-throughput identification of the sensitivities of an Escherichia coli ΔrecA mutant strain to various chemical compounds
复制标题
DOI：
10.1038/s41429-019-0160-5
复制
发表时间：
2019-02
期刊：
影响因子：
--
通讯作者：
Tomoya Maeda;Takaaki Horinouchi;Natsue Sakata;Akio Sakai;C. Furusawa
中科院分区：
文献类型：
--
作者：
Tomoya Maeda;Takaaki Horinouchi;Natsue Sakata;Akio Sakai;C. Furusawa
抗生素耐药性被认为是对公共卫生的全球威胁。适应性耐药突变和通过水平基因转移获得耐药基因已知在抗生素治疗期间由RecA依赖性SOS反应促进，使得RecA抑制剂有希望用于预防抗生素耐药性。然而，RecA失活对抗生素敏感性的影响仍不清楚。因此，在本研究中，我们进行了高通量筛选，以确定217种化学物质的最低抑菌浓度（MIC），包括抗生素和未知药物作用的有毒化学物质，在野生型MDS 42和Δ recA突变株的大肠杆菌。Δ recA突变体对DNA损伤剂、DNA复制抑制剂和铬酸盐胁迫以及其他化学物质如S-（2-氨乙基）-l-半胱氨酸、l-组氨酸、钌红、d-青霉胺、羰基氰3-氯苯腙（CCCP）、浅蓝菌素和l-半胱氨酸的敏感性增加。基因芯片分析进一步表明，与野生型相比，Δ recA突变体中编码氮同化调节因子的glnK、nac和glnLG以及编码铵转运蛋白的amtB的表达量较低。这些结果表明，ΔrecAmutation不仅影响SOS反应，而且影响氨基酸代谢。
Antibiotic resistance is considered a global threat to public health. Adaptive resistance mutations and the acquisition of resistance genes by horizontal gene transfer are known to be facilitated by the RecA-dependent SOS response during antibiotic treatment, making RecA inhibitors promising agents for the prevention of antibiotic resistance. However, the impact of RecA inactivation on antibiotic sensitivities remains unclear. Therefore, in this study, we performed high-throughput screening to determine the minimum inhibitory concentrations (MICs) of 217 chemicals, including both antibiotics and toxic chemicals of unknown drug action, in the wild-type MDS42 and the ΔrecAmutant strains ofEscherichia coli. The ΔrecAmutant showed increased sensitivity to DNA-damaging agents, DNA replication inhibitors, and chromate stress, as well as to other chemicals, such asS-(2-aminoethyl)-l-cysteine,l-histidine, ruthenium red,d-penicillamine, carbonyl cyanide 3-chlorophenylhydrazone (CCCP), cerulenin, andl-cysteine. Microarray analysis showed further that the ΔrecAmutant had lower expressions ofglnK,nac, andglnLG, which encode nitrogen assimilation regulators, as well asamtB, which encodes an ammonium transporter, compared with the wild type. These findings suggest that the ΔrecAmutation affects not only the SOS response but also amino acid metabolism.