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Phosphoregulated orthogonal signal transduction in mammalian cells.

  • Scheller L Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, CH-4058, Basel, Switzerland.
  • Schmollack M Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, CH-4058, Basel, Switzerland.
  • Bertschi A Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, CH-4058, Basel, Switzerland.
  • Mansouri M Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, CH-4058, Basel, Switzerland.
  • Saxena P Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, CH-4058, Basel, Switzerland.
  • Fussenegger M Department of Biosystems Science and Engineering, ETH Zurich, Mattenstrasse 26, CH-4058, Basel, Switzerland. martin.fussenegger@bsse.ethz.ch.
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  • 2020-06-20
Published in:
  • Nature communications. - 2020
Adipose Tissue
Animals
Bacterial Proteins
Female
Gene Expression Regulation
HEK293 Cells
Histidine
Histidine Kinase
Humans
Induced Pluripotent Stem Cells
Mesenchymal Stem Cells
Middle Aged
Nanotechnology
Phosphorylation
Protein Domains
Protein Multimerization
Protein Processing, Post-Translational
Signal Transduction
Synthetic Biology
Transcription Factors
English Orthogonal tools for controlling protein function by post-translational modifications open up new possibilities for protein circuit engineering in synthetic biology. Phosphoregulation is a key mechanism of signal processing in all kingdoms of life, but tools to control the involved processes are very limited. Here, we repurpose components of bacterial two-component systems (TCSs) for chemically induced phosphotransfer in mammalian cells. TCSs are the most abundant multi-component signal-processing units in bacteria, but are not found in the animal kingdom. The presented phosphoregulated orthogonal signal transduction (POST) system uses induced nanobody dimerization to regulate the trans-autophosphorylation activity of engineered histidine kinases. Engineered response regulators use the phosphohistidine residue as a substrate to autophosphorylate an aspartate residue, inducing their own homodimerization. We verify this approach by demonstrating control of gene expression with engineered, dimerization-dependent transcription factors and propose a phosphoregulated relay system of protein dimerization as a basic building block for next-generation protein circuits.
Language
  • English
Open access status
gold
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Persistent URL
https://sonar.ch/global/documents/158797
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