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Phosphate sink containing two-component signaling systems as tunable threshold devices.

Munia Amin1, Varun B Kothamachu2, Elisenda Feliu3

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Summary

Synthetic biology can reengineer signaling circuits using a histidine kinase (HK) and two response regulators (RR). This motif enables tunable, switch-like signal processing and rapid signal termination for threshold devices.

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Area of Science:

  • Synthetic biology
  • Molecular systems biology
  • Biochemical signaling

Background:

  • Reengineering biological signaling circuits is challenging due to limited understanding of system dynamics.
  • Bacterial two-component signaling systems, based on histidine kinase (HK) and response regulator (RR) proteins, offer a promising platform for synthetic biology.
  • A specific motif involves one HK reversibly phosphorylating two RRs, impacting signal processing.

Purpose of the Study:

  • To explore the signal-response relationship of a one HK-two RR motif.
  • To identify conditions for sigmoidal signal-response relationships and control parameters.
  • To provide a basis for developing threshold devices in synthetic biology.

Main Methods:

  • Mathematical modeling of the one HK-two RR motif.
  • In vitro reconstitution of the motif from Sinorhizobium meliloti.
  • Experimental measurement of signal-response curves and sigmoidality control.

Main Results:

  • The one HK-two RR motif enables rapid signal termination via a phosphate sink RR.
  • This motif implements a sigmoidal signal-response relationship, with identified mathematical conditions.
  • Sigmoidality is experimentally tunable via the sink RR and an auxiliary protein interacting with the HK.

Conclusions:

  • The one HK-two RR motif allows for tunable, switch-like signal processing in bacteria and yeast.
  • This motif provides a robust foundation for designing threshold devices in synthetic biology.
  • Understanding and engineering these signaling motifs advance synthetic biology applications.