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Engineering DNA recognition and allosteric response properties of TetR family proteins by using a module-swapping strategy

The development of synthetic biological systems requires modular biomolecular components to flexibly alter response pathways. In previous studies, we have established a module-swapping design principle to engineer allosteric response and DNA recognition properties among regulators in the LacI family...

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Autores principales: Dimas, Rey P, Jordan, Benjamin R, Jiang, Xian-Li, Martini, Catherine, Glavy, Joseph S, Patterson, Dustin P, Morcos, Faruck, Chan, Clement T Y
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6895282/
https://www.ncbi.nlm.nih.gov/pubmed/31392336
http://dx.doi.org/10.1093/nar/gkz666
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author Dimas, Rey P
Jordan, Benjamin R
Jiang, Xian-Li
Martini, Catherine
Glavy, Joseph S
Patterson, Dustin P
Morcos, Faruck
Chan, Clement T Y
author_facet Dimas, Rey P
Jordan, Benjamin R
Jiang, Xian-Li
Martini, Catherine
Glavy, Joseph S
Patterson, Dustin P
Morcos, Faruck
Chan, Clement T Y
author_sort Dimas, Rey P
collection PubMed
description The development of synthetic biological systems requires modular biomolecular components to flexibly alter response pathways. In previous studies, we have established a module-swapping design principle to engineer allosteric response and DNA recognition properties among regulators in the LacI family, in which the engineered regulators served as effective components for implementing new cellular behavior. Here we introduced this protein engineering strategy to two regulators in the TetR family: TetR (UniProt Accession ID: P04483) and MphR (Q9EVJ6). The TetR DNA-binding module and the MphR ligand-binding module were used to create the TetR-MphR. This resulting hybrid regulator possesses DNA-binding properties of TetR and ligand response properties of MphR, which is able to control gene expression in response to a molecular signal in cells. Furthermore, we studied molecular interactions between the TetR DNA-binding module and MphR ligand-binding module by using mutant analysis. Together, we demonstrated that TetR family regulators contain discrete and functional modules that can be used to build biological components with novel properties. This work highlights the utility of rational design as a means of creating modular parts for cell engineering and introduces new possibilities in rewiring cellular response pathways.
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spelling pubmed-68952822019-12-11 Engineering DNA recognition and allosteric response properties of TetR family proteins by using a module-swapping strategy Dimas, Rey P Jordan, Benjamin R Jiang, Xian-Li Martini, Catherine Glavy, Joseph S Patterson, Dustin P Morcos, Faruck Chan, Clement T Y Nucleic Acids Res Synthetic Biology and Bioengineering The development of synthetic biological systems requires modular biomolecular components to flexibly alter response pathways. In previous studies, we have established a module-swapping design principle to engineer allosteric response and DNA recognition properties among regulators in the LacI family, in which the engineered regulators served as effective components for implementing new cellular behavior. Here we introduced this protein engineering strategy to two regulators in the TetR family: TetR (UniProt Accession ID: P04483) and MphR (Q9EVJ6). The TetR DNA-binding module and the MphR ligand-binding module were used to create the TetR-MphR. This resulting hybrid regulator possesses DNA-binding properties of TetR and ligand response properties of MphR, which is able to control gene expression in response to a molecular signal in cells. Furthermore, we studied molecular interactions between the TetR DNA-binding module and MphR ligand-binding module by using mutant analysis. Together, we demonstrated that TetR family regulators contain discrete and functional modules that can be used to build biological components with novel properties. This work highlights the utility of rational design as a means of creating modular parts for cell engineering and introduces new possibilities in rewiring cellular response pathways. Oxford University Press 2019-09-19 2019-08-08 /pmc/articles/PMC6895282/ /pubmed/31392336 http://dx.doi.org/10.1093/nar/gkz666 Text en © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Synthetic Biology and Bioengineering
Dimas, Rey P
Jordan, Benjamin R
Jiang, Xian-Li
Martini, Catherine
Glavy, Joseph S
Patterson, Dustin P
Morcos, Faruck
Chan, Clement T Y
Engineering DNA recognition and allosteric response properties of TetR family proteins by using a module-swapping strategy
title Engineering DNA recognition and allosteric response properties of TetR family proteins by using a module-swapping strategy
title_full Engineering DNA recognition and allosteric response properties of TetR family proteins by using a module-swapping strategy
title_fullStr Engineering DNA recognition and allosteric response properties of TetR family proteins by using a module-swapping strategy
title_full_unstemmed Engineering DNA recognition and allosteric response properties of TetR family proteins by using a module-swapping strategy
title_short Engineering DNA recognition and allosteric response properties of TetR family proteins by using a module-swapping strategy
title_sort engineering dna recognition and allosteric response properties of tetr family proteins by using a module-swapping strategy
topic Synthetic Biology and Bioengineering
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6895282/
https://www.ncbi.nlm.nih.gov/pubmed/31392336
http://dx.doi.org/10.1093/nar/gkz666
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