Ryu Min-Hyung, Fomicheva Anastasia, O'Neal Lindsey, Alexandre Gladys, Gomelsky Mark
Department of Molecular Biology, University of Wyoming, 1000 E. University Ave, Laramie, WY, 82071, USA.
Current address: Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
Methods Mol Biol. 2017;1657:169-186. doi: 10.1007/978-1-4939-7240-1_14.
Signaling pathways involving second messenger c-di-GMP regulate various aspects of bacterial physiology and behavior. We describe the use of a red light-activated diguanylate cyclase (c-di-GMP synthase) and a blue light-activated c-di-GMP phosphodiesterase (hydrolase) for manipulating intracellular c-di-GMP levels in bacterial cells. We illustrate the application of these enzymes in regulating several c-di-GMP-dependent phenotypes, i.e., motility and biofilm phenotypes in E. coli and chemotactic behavior in the alphaproteobacterium Azospirillum brasilense. We expect these light-activated enzymes to be also useful in regulating c-di-GMP-dependent processes occurring at the fast timescale, for spatial control of bacterial populations, as well as for analyzing c-di-GMP-dependent phenomena at the single-cell level.
涉及第二信使环二鸟苷酸(c-di-GMP)的信号通路调节细菌生理学和行为的各个方面。我们描述了使用红光激活的二鸟苷酸环化酶(c-di-GMP合酶)和蓝光激活的c-di-GMP磷酸二酯酶(水解酶)来操纵细菌细胞内的c-di-GMP水平。我们阐述了这些酶在调节几种c-di-GMP依赖性表型中的应用,即大肠杆菌中的运动性和生物膜表型,以及巴西固氮螺菌中的趋化行为。我们期望这些光激活酶在调节快速时间尺度上发生的c-di-GMP依赖性过程、对细菌群体进行空间控制以及在单细胞水平分析c-di-GMP依赖性现象方面也有用处。