Department of Ecophysiology, Max Planck Institute for Terrestrial Microbiology, Karl-von-Frisch-Str. 10, 35043, Marburg, Germany.
Institute of Molecular Oncology, Member of the German Center for Lung Research (DZL), Philipps-University, Hans-Meerwein-Str. 3, 35043, Marburg, Germany.
Nat Commun. 2020 May 13;11(1):2381. doi: 10.1038/s41467-020-16169-w.
Many bacteria employ a type III secretion system (T3SS) injectisome to translocate proteins into eukaryotic host cells. Although the T3SS can efficiently export heterologous cargo proteins, a lack of target cell specificity currently limits its application in biotechnology and healthcare. In this study, we exploit the dynamic nature of the T3SS to govern its activity. Using optogenetic interaction switches to control the availability of the dynamic cytosolic T3SS component SctQ, T3SS-dependent effector secretion can be regulated by light. The resulting system, LITESEC-T3SS (Light-induced translocation of effectors through sequestration of endogenous components of the T3SS), allows rapid, specific, and reversible activation or deactivation of the T3SS upon illumination. We demonstrate the light-regulated translocation of heterologous reporter proteins, and induction of apoptosis in cultured eukaryotic cells. LITESEC-T3SS constitutes a new method to control protein secretion and translocation into eukaryotic host cells with unparalleled spatial and temporal resolution.
许多细菌利用一种 III 型分泌系统（T3SS）注射器将蛋白质转运到真核宿主细胞中。尽管 T3SS 可以有效地将异源货物蛋白输出，但缺乏靶细胞特异性目前限制了其在生物技术和医疗保健中的应用。在这项研究中，我们利用 T3SS 的动态性质来控制其活性。使用光遗传学相互作用开关来控制动态细胞质 T3SS 成分 SctQ 的可用性，可以通过光来调节 T3SS 依赖性效应物的分泌。由此产生的系统，LITESEC-T3SS（通过隔离 T3SS 的内源性成分来诱导效应物的易位）可以在光照下快速、特异性和可逆地激活或失活 T3SS。我们证明了异源报告蛋白的光调控易位，并诱导培养的真核细胞凋亡。LITESEC-T3SS 构成了一种新的方法，可以在无与伦比的时空分辨率下控制蛋白质分泌和转运到真核宿主细胞中。
