Hefei National Laboratory for Physical Sciences at the Microscale; Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei, 230026, China.
CAS Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, China.
ACS Synth Biol. 2021 Jun 18;10(6):1520-1530. doi: 10.1021/acssynbio.1c00075. Epub 2021 Jun 2.
Bacterial pathogens operate by tightly controlling the pathogenicity to facilitate invasion and survival in host. While small molecule inducers can be designed to modulate pathogenicity to perform studies of pathogen-host interaction, these approaches, due to the diffusion property of chemicals, may have unintended, or pleiotropic effects that can impose limitations on their use. By contrast, light provides superior spatial and temporal resolution. Here, using optogenetics we reengineered GacS of the opportunistic pathogen , signal transduction protein of the global regulatory Gac/Rsm cascade which is of central importance for the regulation of infection factors. The resultant protein (termed YGS24) displayed significant light-dependent activity of GacS kinases in . When introduced in the host systems, YGS24 stimulated the pathogenicity of the strain PAO1 in a brain-heart infusion and of another strain, PA14, in slow killing media progressively upon blue-light exposure. This optogenetic system provides an accessible way to spatiotemporally control bacterial pathogenicity in defined hosts, even specific tissues, to develop new pathogenesis systems, which may in turn expedite development of innovative therapeutics.
细菌病原体通过严格控制致病性来促进在宿主中的入侵和生存。虽然小分子诱导剂可以设计用于调节致病性以进行病原体-宿主相互作用的研究，但由于化学物质的扩散性质，这些方法可能会产生意外的或多效性的影响，从而限制了它们的使用。相比之下，光提供了更高的空间和时间分辨率。在这里，我们使用光遗传学重新设计了机会性病原体 中的 GacS，即全局调控 Gac/Rsm 级联的信号转导蛋白，它对感染因子的调控至关重要。所得蛋白（称为 YGS24）在 中显示出 GacS 激酶的显著光依赖性活性。当引入宿主系统时，YGS24 在脑心灌注中刺激 菌株 PAO1 的致病性，并且在暴露于蓝光时，在慢杀培养基中逐渐刺激另一种菌株 PA14 的致病性。这种光遗传学系统为在特定宿主中甚至特定组织中时空控制细菌致病性提供了一种可行的方法，以开发新的发病机制系统，从而可能加速创新疗法的发展。
