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Artificial signaling in mammalian cells enabled by prokaryotic two-component system

Augmenting live cells with novel signal transduction capabilities is a key objective in genetic engineering and synthetic biology. We showed earlier that two-component signaling pathways could function in mammalian cells, albeit while losing their ligand sensitivity. Here we show how to transduce sm...

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Detalles Bibliográficos
Autores principales: Mazé, Alain, Benenson, Yaakov
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6982536/
https://www.ncbi.nlm.nih.gov/pubmed/31844302
http://dx.doi.org/10.1038/s41589-019-0429-9
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author Mazé, Alain
Benenson, Yaakov
author_facet Mazé, Alain
Benenson, Yaakov
author_sort Mazé, Alain
collection PubMed
description Augmenting live cells with novel signal transduction capabilities is a key objective in genetic engineering and synthetic biology. We showed earlier that two-component signaling pathways could function in mammalian cells, albeit while losing their ligand sensitivity. Here we show how to transduce small molecule ligands in a dose-dependent fashion into gene expression in mammalian cells using two-component signaling machinery. First, we engineer mutually complementing truncated mutants of a histidine kinase unable to dimerize and phosphorylate the response regulator. Next, we fuse these mutants to protein domains capable of ligand-induced dimerization, which restores the phosphoryl transfer in a ligand-dependent manner. Cytoplasmic ligands are transduced by facilitating mutant dimerization in the cytoplasm, while extracellular ligands trigger dimerization at the inner side of a plasma membrane. These findings point to the potential of two-component regulatory systems as enabling tools for orthogonal signaling pathways in mammalian cells.
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spelling pubmed-69825362020-06-16 Artificial signaling in mammalian cells enabled by prokaryotic two-component system Mazé, Alain Benenson, Yaakov Nat Chem Biol Article Augmenting live cells with novel signal transduction capabilities is a key objective in genetic engineering and synthetic biology. We showed earlier that two-component signaling pathways could function in mammalian cells, albeit while losing their ligand sensitivity. Here we show how to transduce small molecule ligands in a dose-dependent fashion into gene expression in mammalian cells using two-component signaling machinery. First, we engineer mutually complementing truncated mutants of a histidine kinase unable to dimerize and phosphorylate the response regulator. Next, we fuse these mutants to protein domains capable of ligand-induced dimerization, which restores the phosphoryl transfer in a ligand-dependent manner. Cytoplasmic ligands are transduced by facilitating mutant dimerization in the cytoplasm, while extracellular ligands trigger dimerization at the inner side of a plasma membrane. These findings point to the potential of two-component regulatory systems as enabling tools for orthogonal signaling pathways in mammalian cells. 2019-12-16 2020-02 /pmc/articles/PMC6982536/ /pubmed/31844302 http://dx.doi.org/10.1038/s41589-019-0429-9 Text en http://www.nature.com/authors/editorial_policies/license.html#terms Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Mazé, Alain
Benenson, Yaakov
Artificial signaling in mammalian cells enabled by prokaryotic two-component system
title Artificial signaling in mammalian cells enabled by prokaryotic two-component system
title_full Artificial signaling in mammalian cells enabled by prokaryotic two-component system
title_fullStr Artificial signaling in mammalian cells enabled by prokaryotic two-component system
title_full_unstemmed Artificial signaling in mammalian cells enabled by prokaryotic two-component system
title_short Artificial signaling in mammalian cells enabled by prokaryotic two-component system
title_sort artificial signaling in mammalian cells enabled by prokaryotic two-component system
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6982536/
https://www.ncbi.nlm.nih.gov/pubmed/31844302
http://dx.doi.org/10.1038/s41589-019-0429-9
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