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Consequences of producing DNA gyrase from a synthetic gyrBA operon in Salmonella enterica serovar Typhimurium

German Pozdeev, View ORCID ProfileAalap Mogre, View ORCID ProfileCharles J Dorman
doi: https://doi.org/10.1101/2020.12.08.416404
German Pozdeev
Department of Microbiology, Moyne Institute of Preventive Medicine, Trinity College Dublin, Dublin 2, Ireland
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Aalap Mogre
Department of Microbiology, Moyne Institute of Preventive Medicine, Trinity College Dublin, Dublin 2, Ireland
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Charles J Dorman
Department of Microbiology, Moyne Institute of Preventive Medicine, Trinity College Dublin, Dublin 2, Ireland
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  • For correspondence: cjdorman@tcd.ie
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Abstract

DNA gyrase is an essential type II topoisomerase that is composed of two subunits, GyrA and GyrB and has an A2B2 structure. Although both subunits are required in equal proportions to form DNA gyrase, the gyrA and gyrB genes that encode them in Salmonella (and in many other bacteria) are at widely separated locations on the chromosome, are under separate transcriptional control and are present in different copy numbers in rapidly growing bacteria (gyrA is near the terminus of chromosome replication while gyrB is near the origin). We generated a synthetic gyrBA operon at the oriC-proximal location of gyrB to test the significance of the gyrase gene position for Salmonella physiology. Producing gyrase from an operon did not alter growth kinetics, cell morphology, competitive fitness index, or sensitivity to some gyrase-inhibiting antibiotics. However, the operon strain had altered DNA supercoiling set points, its SPI-2 virulence genes were expressed at a reduced level and its survival was reduced in macrophage. The gyrB gene could not be deleted from its oriC-proximal location, even in a gyrB merodiploid strain. We discuss the physiological significance of the different gyrA and gyrB gene arrangements found naturally in Salmonella and other bacteria.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted December 08, 2020.
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Consequences of producing DNA gyrase from a synthetic gyrBA operon in Salmonella enterica serovar Typhimurium
German Pozdeev, Aalap Mogre, Charles J Dorman
bioRxiv 2020.12.08.416404; doi: https://doi.org/10.1101/2020.12.08.416404
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Consequences of producing DNA gyrase from a synthetic gyrBA operon in Salmonella enterica serovar Typhimurium
German Pozdeev, Aalap Mogre, Charles J Dorman
bioRxiv 2020.12.08.416404; doi: https://doi.org/10.1101/2020.12.08.416404

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