使用基于 рибо开关的生物传感器识别调节c-di-AMP生产的新型遗传因素在金黄色葡萄球菌中
Igor Kviatkovski1, Qiyun Zhong1, Sanika Vaidya1
1Section of Molecular Microbiology and Centre for Bacterial Resistance Biology, Imperial College London, London, United Kingdom.
mSphere
|September 17, 2024
概括
我们确定了核糖核酸还原酶作为一种新型调节器的循环二-AMP (c-di-AMP) 在黄金葡萄球菌的水平. 减少其表达会增加c-di-AMP,影响抗生素耐药性.
科学领域:
- 细菌生理学和信号传递
- 分子微生物学分子微生物学
- 抗微生物药物耐药性的机制
背景情况:
- 循环二-AMP (c-di-AMP) 是细菌 (包括金黄色葡萄球菌) 中的关键次要信使,调节重要过程,如吸收和抗生素耐药性.
- 细胞c-di-AMP水平通过合成和水解受到严格控制,但新的调节因素在很大程度上是未知的.
- 黄金葡萄球菌 (Staphylococcus aureus) 是一个重要的人类病原体,表现出增强的β-乳酸胺抗性,并增加了c-di-AMP水平.
研究的目的:
- 开发和验证一种生物传感器,用于检测Staphylococcus aureus中的c-di-AMP水平.
- 通过转子子子突变图书馆屏幕识别调节细胞内c-di-AMP度的新型遗传因素.
- 阐明已识别的因素在c-di-AMP平衡中的作用及其与抗生素耐药性的联系.
主要方法:
- 使用 Bacillus subtilis kimA riboswitch 和 yfp 记者基因构建一个 c-di-AMP 生物传感器.
- 使用c-di-AMP生物传感器对 Staphylococcus aureus 转体子突变库进行高通量选.
- 对表现出改变c-di-AMP水平的突变物进行基因分析,包括测序和基因表达研究.
主要成果:
- 生物传感器成功地检测到了Staphylococcus aureus中的差异c-di-AMP水平.
- 选验证了已知的调节器 (gdpP,cdaR) 并确定了具有增加c-di-AMP的新突变.
- 在nrdIEF操作子上游的突变,编码核糖核酸还原酶,导致nrdIEF表达率下降和c-di-AMP水平升高.
- 这种调节被发现依赖于Rsh, (p) pGpp的合成酶,GdpP的抑制剂.
结论:
- 建立了一个强大的生物传感器系统,用于在金黄色葡萄球菌中检测c-di-AMP,可适应其他细菌物种.
- 核酸缩酶 (NrdIEF) 被确定为一种新型的负调节剂,用于 Staphylococcus aureus 中的 c-di-AMP 生产.
- 了解c-di-AMP调节,特别是其与MRSA等病原体抗生素耐药性的联系,对于开发新的治疗策略至关重要.
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