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Reconstitution and Coupling of DNA Replication and Segregation in a Biomimetic System

A biomimetic system capable of replication and segregation of genetic material constitutes an essential component for the future design of a minimal synthetic cell. Here we have used the simple T7 bacteriophage system and the plasmid‐derived ParMRC system to establish in vitro DNA replication and DN...

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Autores principales: Hürtgen, Daniel, Mascarenhas, Judita, Heymann, Michael, Murray, Seán M., Schwille, Petra, Sourjik, Victor
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6899551/
https://www.ncbi.nlm.nih.gov/pubmed/31344304
http://dx.doi.org/10.1002/cbic.201900299
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author Hürtgen, Daniel
Mascarenhas, Judita
Heymann, Michael
Murray, Seán M.
Schwille, Petra
Sourjik, Victor
author_facet Hürtgen, Daniel
Mascarenhas, Judita
Heymann, Michael
Murray, Seán M.
Schwille, Petra
Sourjik, Victor
author_sort Hürtgen, Daniel
collection PubMed
description A biomimetic system capable of replication and segregation of genetic material constitutes an essential component for the future design of a minimal synthetic cell. Here we have used the simple T7 bacteriophage system and the plasmid‐derived ParMRC system to establish in vitro DNA replication and DNA segregation, respectively. These processes were incorporated into biomimetic compartments providing an enclosed reaction space. The functional lifetime of the encapsulated segregation system could be prolonged by equipping it with ATP‐regenerating and oxygen‐scavenging systems. Finally, we showed that DNA replication and segregation processes could be coupled in vitro by using condensed DNA nanoparticles resulting from DNA replication. ParM spindles extended over tens of micrometers and could thus be used for segregation in compartments that are significantly longer than bacterial cell size. Overall, this work demonstrates the successful bottom‐up assembly and coupling of molecular machines that mediate replication and segregation, thus providing an important step towards the development of a fully functional minimal cell.
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spelling pubmed-68995512019-12-19 Reconstitution and Coupling of DNA Replication and Segregation in a Biomimetic System Hürtgen, Daniel Mascarenhas, Judita Heymann, Michael Murray, Seán M. Schwille, Petra Sourjik, Victor Chembiochem Full Papers A biomimetic system capable of replication and segregation of genetic material constitutes an essential component for the future design of a minimal synthetic cell. Here we have used the simple T7 bacteriophage system and the plasmid‐derived ParMRC system to establish in vitro DNA replication and DNA segregation, respectively. These processes were incorporated into biomimetic compartments providing an enclosed reaction space. The functional lifetime of the encapsulated segregation system could be prolonged by equipping it with ATP‐regenerating and oxygen‐scavenging systems. Finally, we showed that DNA replication and segregation processes could be coupled in vitro by using condensed DNA nanoparticles resulting from DNA replication. ParM spindles extended over tens of micrometers and could thus be used for segregation in compartments that are significantly longer than bacterial cell size. Overall, this work demonstrates the successful bottom‐up assembly and coupling of molecular machines that mediate replication and segregation, thus providing an important step towards the development of a fully functional minimal cell. John Wiley and Sons Inc. 2019-08-28 2019-10-15 /pmc/articles/PMC6899551/ /pubmed/31344304 http://dx.doi.org/10.1002/cbic.201900299 Text en © 2019 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Full Papers
Hürtgen, Daniel
Mascarenhas, Judita
Heymann, Michael
Murray, Seán M.
Schwille, Petra
Sourjik, Victor
Reconstitution and Coupling of DNA Replication and Segregation in a Biomimetic System
title Reconstitution and Coupling of DNA Replication and Segregation in a Biomimetic System
title_full Reconstitution and Coupling of DNA Replication and Segregation in a Biomimetic System
title_fullStr Reconstitution and Coupling of DNA Replication and Segregation in a Biomimetic System
title_full_unstemmed Reconstitution and Coupling of DNA Replication and Segregation in a Biomimetic System
title_short Reconstitution and Coupling of DNA Replication and Segregation in a Biomimetic System
title_sort reconstitution and coupling of dna replication and segregation in a biomimetic system
topic Full Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6899551/
https://www.ncbi.nlm.nih.gov/pubmed/31344304
http://dx.doi.org/10.1002/cbic.201900299
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