Re-engineering the two-component systems as light-regulated in Escherichia coli |
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| Authors: | Siya Ma Siwei Luo Li Wu Zhi Liang Jia-Rui Wu |
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| Affiliation: | 1.Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences,University of Science and Technology of China,Hefei,China;2.Key Laboratory of Systems Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences,Chinese Academy of Sciences,Shanghai,China;3.School of Life Science and Technology,ShanghaiTech University,Shanghai,China |
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| Abstract: | Bacteria live in environments with dynamic changes. To sense and respond to different external stimuli, bacteria make use of various sensor-response circuits, called two-component systems (TCSs). A TCS comprises a histidine protein kinase (HK) sensing environmental stimuli and a response regulator protein (RR) regulating downstream genes. The two components are coupled via a phosphorylation control mechanism. In a recent study, we adopted an optogenetics approach to re-engineer the sensor HKs in Escherichia coli as a light-sensing fusion protein. We constructed a light-controllable HK by replacing the original signal-specific sensing domain of HK with the light-sensing domain of Cph1 from Cyanobacteria Synechocystis, so that HK can be investigated by red light. Here, we extended the study to other 16 HK-RR TCSs and constructed a library of light-responsible HK-Cph1 chimeras. By taking the NarX-NarL system as an example, we demonstrated the light responsiveness of the constructed chimera and investigated the frequency response of the NarX-NarL system. The constructed library serves as a toolkit for future TCS study using optogenetics approach. |
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