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Genomic signatures of experimental adaptation to antimicrobial peptides in Staphylococcus aureus

Paul R. Johnston, Adam J. Dobson, Jens Rolff
doi: https://doi.org/10.1101/023549
Paul R. Johnston
1Freie Universität Berlin, Königin-Luise-Straße 1-3, 14195 Berlin, Germany
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  • For correspondence: paul.johnston@fu-berlin.de
Adam J. Dobson
2University College London, Darwin Building, Gower Street, London, WC1E 6BT, UK
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Jens Rolff
1Freie Universität Berlin, Königin-Luise-Straße 1-3, 14195 Berlin, Germany
3Berlin-Brandenburg Institute of Advanced Biodiversity Research (BBIB), Altensteinstraße 6, 14195, Berlin, Germany
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Abstract

Objectives The evolution of resistance against antimicrobial peptides has long been considered unlikely due to their mechanism of action, yet experimental selection with AMPs results in rapid evolution of resistance in several species of bacteria. Although numerous studies have utilized mutant screens to identify loci that determine AMP susceptibility, there is a dearth of data concerning the genomic changes which accompany experimental evolution of AMP resistance.

Methods Using genome re-sequencing we analysed the mutations which arise during experimental evolution of resistance to the cationic AMPs iseganan, melittin and pexiganan, as well as to a combination of melittin and pexiganan, or to the aminoglycoside antibiotic streptomycin.

Results Analysis of 17 independently replicated Staphylococcus aureus selection lines, including unselected controls, showed that each AMP selected for mutations at distinct loci. We identify mutations in genes involved in the synthesis and maintenance of the cell envelope. This includes genes previously identified from mutant screens for AMP resistance, and genes involved in the response to AMPs and cell-wall-active antibiotics. Furthermore, transposon insertion mutants were used to verify that a number of the identified genes are directly involved in determining AMP susceptibility.

Conclusions Strains selected for AMP resistance under controlled experimental evolution displayed consistent AMP-specific mutations in genes which determine AMP susceptibility. This suggests that different routes to evolve resistance are favored within a controlled genetic background.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted July 30, 2015.
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Genomic signatures of experimental adaptation to antimicrobial peptides in Staphylococcus aureus
Paul R. Johnston, Adam J. Dobson, Jens Rolff
bioRxiv 023549; doi: https://doi.org/10.1101/023549
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Genomic signatures of experimental adaptation to antimicrobial peptides in Staphylococcus aureus
Paul R. Johnston, Adam J. Dobson, Jens Rolff
bioRxiv 023549; doi: https://doi.org/10.1101/023549

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