Extracytoplasmic Function σ Factors Can Be Implemented as Robust Heterologous Genetic Switches in Bacillus subtilis

Research output: Contribution to journal › Research article › Contributed › peer-review

Contributors

  • Daniela Pinto - , Institute of Microbiology, TUD Dresden University of Technology (Author)
  • Franziska Dürr - , Institute of Microbiology, TUD Dresden University of Technology (Author)
  • Friederike Froriep - , Ludwig Maximilian University of Munich (Author)
  • Dayane Araújo - , Ludwig Maximilian University of Munich (Author)
  • Qiang Liu - , Ludwig Maximilian University of Munich (Author)
  • Thorsten Mascher - , Institute of Microbiology, TUD Dresden University of Technology (Author)

Abstract

In bacteria, the promoter specificity of RNA polymerase is determined by interchangeable σ subunits. Extracytoplasmic function σ factors (ECFs) form the largest and most diverse family of alternative σ factors, and their suitability for constructing genetic switches and circuits was already demonstrated. However, a systematic study on how genetically determined perturbations affect the behavior of these switches is still lacking, which impairs our ability to predict their behavior in complex circuitry. Here, we implemented four ECF switches in Bacillus subtilis and comprehensively characterized their robustness toward genetic perturbations, including changes in copy number, protein stability, or antisense transcription. All switches show characteristic dose-response behavior that varies depending on the individual ECF-promoter pair. Most perturbations had performance costs. Although some general design rules could be derived, a detailed characterization of each ECF switch before implementation is recommended to understand and thereby accommodate its individual behavior.

Details

Original languageEnglish
Pages (from-to)380-390
Number of pages11
JournaliScience
Volume13
Publication statusPublished - 29 Mar 2019
Peer-reviewedYes

External IDs

PubMedCentral PMC6426705
Scopus 85066291392

Keywords

Library keywords