A fungicide-responsive kinase as a tool for synthetic cell fate regulation

Nucleic Acids Res. 2015 Aug 18;43(14):7162-70. doi: 10.1093/nar/gkv678. Epub 2015 Jul 2.

Abstract

Engineered biological systems that precisely execute defined tasks have major potential for medicine and biotechnology. For instance, gene- or cell-based therapies targeting pathogenic cells may replace time- and resource-intensive drug development. Engineering signal transduction systems is a promising, yet presently underexplored approach. Here, we exploit a fungicide-responsive heterologous histidine kinase for pathway engineering and synthetic cell fate regulation in the budding yeast Saccharomyces cerevisiae. Rewiring the osmoregulatory Hog1 MAPK signalling system generates yeast cells programmed to execute three different tasks. First, a synthetic negative feedback loop implemented by employing the fungicide-responsive kinase and a fungicide-resistant derivative reshapes the Hog1 activation profile, demonstrating how signalling dynamics can be engineered. Second, combinatorial integration of different genetic parts including the histidine kinases, a pathway activator and chemically regulated promoters enables control of yeast growth and/or gene expression in a two-input Boolean logic manner. Finally, we implemented a genetic 'suicide attack' system, in which engineered cells eliminate target cells and themselves in a specific and controllable manner. Taken together, fungicide-responsive kinases can be applied in different constellations to engineer signalling behaviour. Sensitizing engineered cells to existing chemicals may be generally useful for future medical and biotechnological applications.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antifungal Agents / pharmacology*
  • Cell Engineering / methods*
  • DNA-Binding Proteins / metabolism
  • Dioxoles / pharmacology
  • Feedback, Physiological
  • Gene Expression Regulation*
  • Histidine Kinase
  • MAP Kinase Signaling System / drug effects
  • MAP Kinase Signaling System / genetics*
  • Mitogen-Activated Protein Kinases / metabolism*
  • Protein Kinases / genetics
  • Pyrroles / pharmacology
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Transcription Factors / metabolism

Substances

  • Antifungal Agents
  • DNA-Binding Proteins
  • Dioxoles
  • Pyrroles
  • SKN7 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors
  • Protein Kinases
  • HOG1 protein, S cerevisiae
  • Mitogen-Activated Protein Kinases
  • Histidine Kinase
  • fludioxonil