Repurposing Synechocystis PCC6803 UirS-UirR as a UV-Violet/Green Photoreversible Transcriptional Regulatory Tool in E. coli.
We have previously engineered green/red and red/far red photoreversible E. coli phytochrome and cyanobacteriochrome (CBCR) two-component systems (TCSs) and utilized them to program tailor-made gene expression signals for gene circuit characterization. Here, we transport the UV-violet/green photoreversible CBCR TCS UirS-UirR from Synechocystis PCC6803 to E. coli. We demonstrate that the promoter of the small RNA csiR1, previously shown to be activated by inorganic carbon stress, is a UirS-UirR output. Additionally, in contrast to a recently proposed sequestration model, we show that the sensor histidine kinase UirS phosphorylates the response regulator UirR to activate PcsiR1 transcription in response to UV-violet light. Finally, we measure changes in UirS-UirR output minutes after a change in light input and exploit these rapid dynamics to program a challenging gene expression signal with high predictability. UirS-UirR is the first engineered transcriptional regulatory tool activated exclusively by UV-violet light, and the most blue shifted photoreversible transcriptional regulatory tool.
Department of Bioengineering, ‡Department of Biosciences, Rice University , 6100 Main Street, Houston, Texas 77005, United States.
Prabha
P
Ramakrishnan
Department of Bioengineering, ‡Department of Biosciences, Rice University , 6100 Main Street, Houston, Texas 77005, United States.
Jeffrey J
JJ
Tabor
D001426:Bacterial Proteins / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005818:Genetic Engineering / Q000379:methods*; D062308:Optogenetics / Q000379:methods*; D010766:Phosphorylation; D046939:Synechocystis / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic; D014466:Ultraviolet Rays
D001426:Bacterial Proteins
UV-violet light; bacterial two-component system; light-regulated gene expression; optogenetics