In bacteria, cyclic adenosine monophosphate (cAMP) signaling plays an essential regulatory role whose modulation via optogenetic tools would provide researchers an immense opportunity to control biological processes simply by illumination. The cAMP signaling in bacteria is a complex network of regulatory pathways, which utilizes distinct proteomic resources under different nutrient environments. We established an optogenetic modulation of cAMP and studied important cellular process of growth, biofilm formation, and virulence in the model bacterium E. coli using a light-gated adenylate cyclase (LgAC) from Beggiatoa sp. Blue light-induced activation of LgAC elevated the cAMP level in a blue light-dependent manner in E. coli. Quantitative proteomics revealed a decrease in the level of certain proteins governing growth (PTS, Adk, AckA, GlnA, and EFP), biofilm formation (IhfA, flagellin, YajQ, YeaG, and HlfC), and virulence (ClpP, YebC, KatE, BtuE, and Zur) in E. coli cells expressing LgAC upon blue light illumination. This optogenetic modulation of cAMP would be useful for deciphering cAMP-associated host–pathogen signaling of bacterial systems. Proteome knowledge established by this research work would also be useful for the scientific community while adapting LgAC-based optogenetic modulation for studying other relevant cAMP-driven bacterial physiology (e.g., energy metabolism). The systematic utilization of the established method and more extensively designed experiments regarding bacterial growth, biofilm, survival, and virulence might provide a road map for the identification of new targets for developing novel antibacterial drugs.

中文翻译：

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建立cAMP的光遗传调制，以使用光门控环化酶分析细菌的生长，生物膜形成和毒力途径

在细菌中，环状单磷酸腺苷（cAMP）信号传导起着至关重要的调节作用，其通过光遗传学工具的调节将为研究人员提供一个巨大的机会，可以通过照明简单地控制生物过程。细菌中的cAMP信号传导是一个复杂的调节通路网络，该通路利用不同营养环境下的独特蛋白质组资源。我们建立了cAMP的光遗传学调制，并使用Beggiatoa sp。的光门控腺苷酸环化酶（LgAC）研究了模型细菌大肠杆菌中重要的细胞生长，生物膜形成和毒力过程。LgAC的蓝色光诱导的活化的升高cAMP水平在蓝色光依赖性大肠杆菌。定量蛋白质组学揭示了控制生长的某些蛋白质（PTS，Adk，AckA，GlnA和EFP），生物膜形成（IhfA，鞭毛蛋白，YajQ，YeaG和HlfC）和毒力（ClpP，YebC，KatE， BtuE和Zur）在大肠杆菌中在蓝光照射下表达LgAC的细胞。cAMP的这种光遗传学调制将有助于破译细菌系统中与cAMP相关的宿主-病原体信号。通过这项研究工作建立的蛋白质组学知识对于科学界也非常有用，同时使基于LgAC的光遗传学调制适应于研究其他相关的cAMP驱动的细菌生理学（例如能量代谢）。关于细菌生长，生物膜，存活和毒力的既定方法的系统利用和更广泛设计的实验，可能为鉴定开发新型抗菌药物的新靶标提供路线图。