Cargando…
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...
Autores principales: | , |
---|---|
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 |
_version_ | 1783491330851209216 |
---|---|
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. |
format | Online Article Text |
id | pubmed-6982536 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT mazealain artificialsignalinginmammaliancellsenabledbyprokaryotictwocomponentsystem AT benensonyaakov artificialsignalinginmammaliancellsenabledbyprokaryotictwocomponentsystem |