Genetically Encoded Chemical Decaging in Living Bacteria

Area of Science:

• Synthetic biology
• Chemical biology
• Microbiology

Background:

• Controlling protein activity within living bacterial cells is crucial for understanding cellular processes and engineering new functions.
• Existing methods like photocaging are often incompatible with the bacterial intracellular environment.
• A robust chemical method is needed for precise protein activation in prokaryotes.

Purpose of the Study:

• To develop a genetically encoded chemical decaging strategy for protein activation in living bacterial cells.
• To establish a general tool for gain-of-function studies in prokaryotic systems.
• To investigate the role of indole in bacterial antibiotic tolerance.

Main Methods:

• Utilized the inverse electron-demand Diels-Alder (iDA) reaction for chemical decaging, ensuring compatibility with the bacterial intracellular environment.
• Applied the iDA strategy for in situ activation of the indole-producing enzyme TnaA in Escherichia coli.
• Constructed an orthogonal and chemically inducible indole production pathway.

Main Results:

• Demonstrated a novel chemical decaging strategy compatible with the intracellular environment of bacteria.
• Successfully activated the TnaA enzyme and established a chemically inducible indole production pathway.
• Revealed a significant role for indole in bacterial antibiotic tolerance.

Conclusions:

• The genetically encoded chemical decaging strategy provides a versatile tool for protein activation in prokaryotes.
• This method allows for precise control over protein function, facilitating gain-of-function studies.
• The study highlights the importance of indole in modulating bacterial antibiotic tolerance.