Wang Xuyang, Li Weizheng, Wang Wenjie, Wang Shiyong, Xu Tao, Chen Jiazhen, Zhang Wenhong
Department of Infectious Diseases, National Medical Center for Infectious Diseases, Shanghai Key Laboratory of Infectious Diseases and Biosafety Emergency Response, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai, China.
Department of Infectious Diseases, Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Front Microbiol. 2021 Dec 23;12:756809. doi: 10.3389/fmicb.2021.756809. eCollection 2021.
Persisters are important reasons for persistent infections, and they can lead to antibiotic treatment failure in patients and consequently chronic infection. small colony variants (SCVs) have been shown to be related to persistent infection. Mutations in the genes of the heme biosynthesis pathway lead to the formation of SCVs. However, the relationship between heme production genes and persister has not been tested. and were knocked out by allelic replacement from strain USA500 separately, and then, the heme deficiency was complemented by overexpression of related genes and the addition of hemin. The stress-related persister assay was conducted. RNA-sequencing was performed to find genes and pathways involved in heme-related persister formation, and relative genes and operons were further knocked out and overexpressed to confirm their role in each process. We found that heme biosynthesis deficiency can lead to decreased persister. After complementing the corresponding genes or hemin, the persister levels could be restored. RNA-seq on knockout strains showed that various metabolic pathways were influenced, such as energy metabolism, amino acid metabolism, carbohydrate metabolism, and membrane transport. Overexpression of and operon could restore USA500∆ persister formation under acid stress. Knocking out operon in USA500∆ could further reduce USA500∆ persister formation under acid and oxidative stress. Heme synthesis has a role in persister formation.
持留菌是持续性感染的重要原因，它们可导致患者抗生素治疗失败并进而引发慢性感染。小菌落变异株（SCVs）已被证明与持续性感染有关。血红素生物合成途径基因的突变会导致SCVs的形成。然而，血红素产生基因与持留菌之间的关系尚未得到验证。分别通过等位基因替换从USA500菌株中敲除 和 ，然后通过相关基因的过表达和添加血红素补充血红素缺乏。进行了与应激相关的持留菌检测。进行RNA测序以寻找参与血红素相关持留菌形成的基因和途径，并进一步敲除和过表达相关基因和操纵子以确认它们在每个过程中的作用。我们发现血红素生物合成缺陷可导致持留菌减少。补充相应基因或血红素后，持留菌水平可恢复。对敲除菌株的RNA测序表明各种代谢途径受到影响，如能量代谢、氨基酸代谢、碳水化合物代谢和膜转运。 和操纵子 的过表达可恢复USA500∆在酸应激下的持留菌形成。在USA500∆中敲除操纵子 可进一步降低USA500∆在酸应激和氧化应激下的持留菌形成。血红素合成在 持留菌形成中起作用。
