Establishing a System for Testing Replication Inhibition of the Vibrio cholerae Secondary Chromosome in Escherichia coli

Schallopp, Nadine; Milbredt, Sarah; Sperlea, Theodor; Kemter, Franziska S.; Bruhn, Matthias; Schindler, Daniel; Waldminghaus, Torsten

doi:10.3390/antibiotics7010003

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Establishing a System for Testing Replication Inhibition of the Vibrio cholerae Secondary Chromosome in Escherichia coli

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LOEWE Center for Synthetic Microbiology-SYNMIKRO, Philipps-Universität Marburg, Marburg 35032, Germany

School of Chemistry, Manchester Institute of Biotechnology, University of Manchester, Manchester M1 7DN, UK

Author to whom correspondence should be addressed.

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N.S., S.M., T.S. and F.S.K. contributed equally to this work.

Academic Editor: Anders Løbner-Olesen

Antibiotics 2018, 7(1), 3; https://doi.org/10.3390/antibiotics7010003

Received: 30 October 2017 / Revised: 5 December 2017 / Accepted: 20 December 2017 / Published: 23 December 2017

(This article belongs to the Special Issue Bacterial DNA Replication and Replication Inhibitors)

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Abstract

Regulators of DNA replication in bacteria are an attractive target for new antibiotics, as not only is replication essential for cell viability, but its underlying mechanisms also differ from those operating in eukaryotes. The genetic information of most bacteria is encoded on a single chromosome, but about 10% of species carry a split genome spanning multiple chromosomes. The best studied bacterium in this context is the human pathogen Vibrio cholerae, with a primary chromosome (Chr1) of 3 M bps, and a secondary one (Chr2) of about 1 M bps. Replication of Chr2 is under control of a unique mechanism, presenting a potential target in the development of V. cholerae-specific antibiotics. A common challenge in such endeavors is whether the effects of candidate chemicals can be focused on specific mechanisms, such as DNA replication. To test the specificity of antimicrobial substances independent of other features of the V. cholerae cell for the replication mechanism of the V. cholerae secondary chromosome, we establish the replication machinery in the heterologous E. coli system. We characterize an E. coli strain in which chromosomal replication is driven by the replication origin of V. cholerae Chr2. Surprisingly, the E. coli ori2 strain was not inhibited by vibrepin, previously found to inhibit ori2-based replication. View Full-Text

Keywords: chromosome engineering; replication initiation; drug development

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MDPI and ACS Style

Schallopp, N.; Milbredt, S.; Sperlea, T.; Kemter, F.S.; Bruhn, M.; Schindler, D.; Waldminghaus, T. Establishing a System for Testing Replication Inhibition of the Vibrio cholerae Secondary Chromosome in Escherichia coli. Antibiotics 2018, 7, 3. https://doi.org/10.3390/antibiotics7010003

AMA Style

Schallopp N, Milbredt S, Sperlea T, Kemter FS, Bruhn M, Schindler D, Waldminghaus T. Establishing a System for Testing Replication Inhibition of the Vibrio cholerae Secondary Chromosome in Escherichia coli. Antibiotics. 2018; 7(1):3. https://doi.org/10.3390/antibiotics7010003

Chicago/Turabian Style

Schallopp, Nadine, Sarah Milbredt, Theodor Sperlea, Franziska S. Kemter, Matthias Bruhn, Daniel Schindler, and Torsten Waldminghaus. 2018. "Establishing a System for Testing Replication Inhibition of the Vibrio cholerae Secondary Chromosome in Escherichia coli" Antibiotics 7, no. 1: 3. https://doi.org/10.3390/antibiotics7010003

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Establishing a System for Testing Replication Inhibition of the Vibrio cholerae Secondary Chromosome in Escherichia coli

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