Construction of a photo-responsive chimeric histidine kinase in Escherichia coli.
Two-component signal transduction systems (TCS), that are also referred to as His to Asp phosphorelay systems, are involved in widespread cellular responses to diverse signals from bacteria to plants. Previously, we succeeded in reconstructing a cyanobacterial photo-perception system in Escherichia coli by employing a CcaS-CcaR two-component system from Nostoc punctiforme. In this study, we have added a photo-responsive ability to ArcB-ArcA (anoxic redox control) TCS of E. coli by fusing a cyanobacterial photoreceptor domain of CcaS with an intracellular histidine kinase (HK) domain of ArcB. For this, we constructed several chimeric HKs between CcaS and ArcB and found that one chimeric HK, named ArcaS9, has a photo-responsive ability. When ArcaS9 was expressed with an ArcA response regulator in E. coli expressing phycocyanobilin (PCB)-producing enzymes, the expression of sdh, a target gene of ArcB-ArcA TCS was regulated in a light-color-dependent manner. Thus we succeeded in endowing E. coli HK with a photo-responsive ability. This provides an insight into how the sensing ability of HK can be manipulated by a chimeric construct.
Laboratory of Molecular Microbiology, Department of Basic Medicinal Sciences, Graduate School of Pharmaceutical Sciences, Nagoya University.
Mayuko
M
Hori
Shyunsuke
S
Oka
Yoshimi
Y
Sugie
Hokuto
H
Ohtsuka
Hirofumi
H
Aiba
D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000528:radiation effects; D015964:Gene Expression Regulation, Bacterial / Q000528:radiation effects*; D000071677:Histidine Kinase / Q000235:genetics / Q000378:metabolism*; D008027:Light; D046937:Nostoc / Q000201:enzymology* / Q000235:genetics; D020283:Photoreceptors, Microbial / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*
D020283:Photoreceptors, Microbial; D011994:Recombinant Proteins; D000071677:Histidine Kinase