Journal
JOURNAL OF BIOSCIENCES
Volume 42, Issue 4, Pages 565-573Publisher
INDIAN ACAD SCIENCES
DOI: 10.1007/s12038-017-9711-8
Keywords
Cph1; histidine kinase; two component systems (TCSs)
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Funding
- National Natural Science Foundation of China [31130034, 31470808]
- Strategic Priority Research Program of the Chinese Academy of Sciences [XDA12000000]
- Ministry of Science and Technology [2012CB917200, 2014CB910600]
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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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