Quadram Institute Bioscience, Norwich Research Park, Norwich NR4 7UQ, UK.
University of East Anglia, Norwich Research Park, Norwich, NR4 7TJ, UK.
Microb Genom. 2021 Dec;7(12). doi: 10.1099/mgen.0.000718.
Trimethoprim and sulfamethoxazole are used commonly together as cotrimoxazole for the treatment of urinary tract and other infections. The evolution of resistance to these and other antibacterials threatens therapeutic options for clinicians. We generated and analysed a chemical-biology-whole-genome data set to predict new targets for antibacterial combinations with trimethoprim and sulfamethoxazole. For this we used a large transposon mutant library in BW25113 where an outward-transcribing inducible promoter was engineered into one end of the transposon. This approach allows regulated expression of adjacent genes in addition to gene inactivation at transposon insertion sites, a methodology that has been called TraDIS-. These chemical genomic data sets identified mechanisms for both reduced and increased susceptibility to trimethoprim and sulfamethoxazole. The data identified that over-expression of FolA reduced trimethoprim susceptibility, a known mechanism for reduced susceptibility. In addition, transposon insertions into the genes , , , , , and increased susceptibility to trimethoprim and likewise for , , , and with sulfamethoxazole, while insertions in , , , , , and increased susceptibility to both these antibiotics. Two of these genes' products, Tdk and IspD, are inhibited by AZT and fosmidomycin respectively, antibiotics that are known to synergise with trimethoprim. Thus, the data identified two known targets and several new target candidates for the development of co-drugs that synergise with trimethoprim, sulfamethoxazole or cotrimoxazole. We demonstrate that the TraDIS- technology can be used to generate information-rich chemical-genomic data sets that can be used for antibacterial development.
甲氧苄啶和磺胺甲噁唑常联合用于治疗尿路感染和其他感染，称为复方新诺明。这些以及其他抗菌药物耐药性的发展，威胁到临床医生的治疗选择。我们生成并分析了一个化学生物学全基因组数据集，以预测与甲氧苄啶和磺胺甲噁唑联合使用的新抗菌药物组合的靶点。为此，我们在 BW25113 中使用了一个大型转座子突变体文库，在该文库中，一个向外转录的诱导启动子被构建到转座子的一端。这种方法除了在转座子插入位点使基因失活之外，还允许相邻基因的调控表达，这种方法被称为 TraDIS-。这些化学基因组数据集确定了对甲氧苄啶和磺胺甲噁唑的敏感性降低和增加的机制。数据表明，FolA 的过表达降低了甲氧苄啶的敏感性，这是一种已知的敏感性降低机制。此外，转座子插入到基因 、 、 、 、 、 和 中，增加了对甲氧苄啶的敏感性，同样，转座子插入到基因 、 、 、 、 、 和 中，增加了对磺胺甲噁唑的敏感性，而转座子插入到基因 、 、 、 、 、 和 中，增加了对这两种抗生素的敏感性。这些基因产物中的两个，Tdk 和 IspD，分别被 AZT 和福米地霉素抑制，这两种抗生素与甲氧苄啶协同作用。因此，这些数据确定了两个已知的靶点和几个新的靶候选物，用于开发与甲氧苄啶、磺胺甲噁唑或复方新诺明协同作用的联合药物。我们证明，TraDIS-技术可用于生成富含信息的化学基因组数据集，可用于抗菌药物的开发。
