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.
- Lindner, F.
- Milne-Davies, B.
- Langenfeld, K.
- Stiewe, T.
- Diepold, A.
Keywords
- Bacterial Proteins/genetics/*metabolism
- Cell Line, Tumor
- Cytosol/metabolism/microbiology
- Eukaryotic Cells/*metabolism/microbiology
- Gene Expression Regulation, Bacterial
- Gram-Negative Bacteria/genetics/*metabolism/physiology
- Humans
- Light
- Microscopy, Fluorescence
- Optogenetics/methods
- Protein Transport/radiation effects
- Spatial Analysis
- Type III Secretion Systems/genetics/*metabolism
- Yersinia enterocolitica/genetics/metabolism/physiology
