Control of the Yeast Mating Pathway by Reconstitution of Functional α-Factor Using Split Intein-Catalyzed Reactions

Area of Science:

• Synthetic biology
• Molecular biology
• Biotechnology

Background:

• Controlling cellular behavior in multicellular organisms and synthetic consortia is challenging due to difficulties in expressing signaling peptides.
• Existing methods for synthetic control are limited by the complexities of heterologous peptide expression.

Purpose of the Study:

• To present a new strategy for reconstituting functional signaling peptides using split intein-mediated reactions.
• To enable precise regulation and coordination of cellular behaviors in synthetic biology applications.

Main Methods:

• Utilized recombinant proteins and split intein-mediated reactions to reconstitute signaling peptides.
• Demonstrated the strategy in Saccharomyces cerevisiae (S. cerevisiae).
• Explored the potential for tailoring peptides and reactions through directed evolution and protein engineering.

Main Results:

• Successfully reconstituted functional signaling peptides capable of eliciting desired cellular responses.
• Showcased a generalizable strategy applicable to various signaling peptides.
• Highlighted the promiscuity of split inteins and the adaptability of the system.

Conclusions:

• The developed strategy offers a versatile platform for implementing synthetic control in diverse biological applications.
• This approach overcomes limitations in heterologous peptide expression for synthetic biology.
• The system is amenable to further optimization through protein engineering for enhanced functionality.