Repurposing Synechocystis PCC6803 UirS–UirR as a UV-Violet/Green Photoreversible Transcriptional Regulatory Tool in E. coli

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• 作者：Prabha Ramakrishnan ; Jeffrey J. Tabor
• 刊名：ACS Synthetic Biology
• 出版年：2016
• 出版时间：July 15, 2016
• 年：2016
• 卷：5
• 期：7
• 页码：733-740
• 全文大小：427K
• 年卷期：0
• ISSN：2161-5063

文摘

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 P_csiR1 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.