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Volume 166, Issue 12

mutations conferring rifampicin-resistance affect growth, stress response and motility in Open Access

Abstract

Rifampicin is a broad-spectrum antibiotic that binds to the bacterial RNA polymerase (RNAP), compromising DNA transcription. Rifampicin resistance is common in several microorganisms and it is typically caused by point mutations in the gene encoding the β subunit of RNA polymerase, . Different mutations are responsible for various levels of rifampicin resistance and for a range of secondary effects. mutations conferring rifampicin resistance have been shown to be responsible for severe effects on transcription, cell fitness, bacterial stress response and virulence. Such effects have never been investigated in the marine pathogen , even though rifampicin-resistant strains of have been isolated previously. Moreover, spontaneous rifampicin-resistant strains of have an important role in conjugation and mutagenesis protocols, with poor consideration of the effects of mutations. In this work, effects on growth, stress response and virulence of were investigated using a set of nine spontaneous rifampicin-resistant derivatives of CMCP6. Three different mutations (Q513K, S522L and H526Y) were identified with varying incidence rates. These three mutant types each showed high resistance to rifampicin [minimal inhibitory concentration (MIC) >800 µg ml], but different secondary effects. The strains carrying the mutation H526Y had a growth advantage in rich medium but had severely reduced salt stress tolerance in the presence of high NaCl concentrations as well as a significant reduction in ethanol stress resistance. Strains possessing the S522L mutation had reduced growth rate and overall biomass accumulation in rich medium. Furthermore, investigation of virulence characteristics demonstrated that all the rifampicin-resistant strains showed compromised motility when compared with the wild-type, but no major effects on exoenzyme production were observed. These findings reveal a wide range of secondary effects of mutations and indicate that rifampicin resistance is not an appropriate selectable marker for studies that aim to investigate phenotypic behaviour in this organism.

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  • Accepted:
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Keyword(s): motility , osmotolerance , rifampicin resistance , rpoB , stress response and Vibrio vulnificus
Funding
This study was supported by the:
  • H2020 Marie Skłodowska-Curie Actions, http://dx.doi.org/10.13039/100010665 (Award 721456)
© 2020 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
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2020-11-13
2024-04-26
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rpoB mutations conferring rifampicin-resistance affect growth, stress response and motility in Vibrio vulnificus
Microbiology 166, 1160 (2020); https://doi.org/10.1099/mic.0.000991
/content/journal/micro/10.1099/mic.0.000991
/content/journal/micro/10.1099/mic.0.000991
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