SynBio and the Boundaries between Functional and Pathogenic RepA-WH1 Bacterial Amyloids

Area of Science:

• Synthetic biology
• Molecular biology
• Microbiology

Background:

• Amyloids, initially linked to diseases, are increasingly recognized as functional protein states across life.
• While most bacterial amyloids are extracellular, intracellular functional amyloids are less understood.
• Distinguishing functional amyloids from pathogenic prions is a key challenge.

Purpose of the Study:

• To review the distinction between functional and pathogenic amyloids from a synthetic biology perspective.
• To explore the RepA-WH1 protein domain, a functional amyloid involved in bacterial plasmid DNA replication.
• To detail the engineering of RepA-WH1 into a synthetic prion using synthetic biology approaches.

Main Methods:

• Utilized protein engineering techniques to modify the RepA-WH1 domain.
• Designed allosteric effectors, including ligands and an optogenetic module, to control RepA-WH1 function.
• Generated an intracellular prion-like agent from RepA-WH1 within bacteria.

Main Results:

• Successfully engineered the RepA-WH1 protein into a synthetic prion-like agent.
• Demonstrated the intracellular cytotoxic potential of the synthetic RepA-WH1 prion.
• Highlighted the role of synthetic biology in creating and controlling amyloid-based agents.

Conclusions:

• Synthetic biology offers powerful tools to manipulate amyloid structures and functions.
• Engineered amyloids, like the synthetic RepA-WH1 prion, can serve as novel antimicrobial agents.
• Further research into synthetic amyloids could bridge the gap between functional and pathogenic protein states.