<italic toggle="yes">In Vivo</italic>-Selected Compensatory Mutations Restore the Fitness Cost of Mosaic <italic toggle="yes">penA</italic> Alleles That Confer Ceftriaxone Resistance in <italic toggle="yes">Neisseria gonorrhoeae</italic>

ABSTRACT Resistance to ceftriaxone in Neisseria gonorrhoeae is mainly conferred by mosaic penA alleles that encode penicillin-binding protein 2 (PBP2) variants with markedly lower rates of acylation by ceftriaxone. To assess the impact of these mosaic penA alleles on gonococcal fitness, we introduce...

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Autores principales:	Leah R. Vincent, Samuel R. Kerr, Yang Tan, Joshua Tomberg, Erica L. Raterman, Julie C. Dunning Hotopp, Magnus Unemo, Robert A. Nicholas, Ann E. Jerse
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	Neisseria gonorrhoeae aconitase antibiotic resistance biological fitness competitive index Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/bf5cc6746a2948e9876c44e5b14525ce
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spelling	oai:doaj.org-article:bf5cc6746a2948e9876c44e5b14525ce2021-11-15T15:53:26Z<italic toggle="yes">In Vivo</italic>-Selected Compensatory Mutations Restore the Fitness Cost of Mosaic <italic toggle="yes">penA</italic> Alleles That Confer Ceftriaxone Resistance in <italic toggle="yes">Neisseria gonorrhoeae</italic>10.1128/mBio.01905-172150-7511https://doaj.org/article/bf5cc6746a2948e9876c44e5b14525ce2018-05-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01905-17https://doaj.org/toc/2150-7511ABSTRACT Resistance to ceftriaxone in Neisseria gonorrhoeae is mainly conferred by mosaic penA alleles that encode penicillin-binding protein 2 (PBP2) variants with markedly lower rates of acylation by ceftriaxone. To assess the impact of these mosaic penA alleles on gonococcal fitness, we introduced the mosaic penA alleles from two ceftriaxone-resistant (Cror) clinical isolates (H041 and F89) into a Cros strain (FA19) by allelic exchange and showed that the resultant Cror mutants were significantly outcompeted by the Cros parent strain in vitro and in a murine infection model. Four Cror compensatory mutants of FA19 penA41 were isolated independently from mice that outcompeted the parent strain both in vitro and in vivo. One of these compensatory mutants (LV41C) displayed a unique growth profile, with rapid log growth followed by a sharp plateau/gradual decline at stationary phase. Genome sequencing of LV41C revealed a mutation (G348D) in the acnB gene encoding the bifunctional aconitate hydratase 2/2 methylisocitrate dehydratase. Introduction of the acnBG348D allele into FA19 penA41 conferred both a growth profile that phenocopied that of LV41C and a fitness advantage, although not as strongly as that exhibited by the original compensatory mutant, suggesting the existence of additional compensatory mutations. The mutant aconitase appears to be a functional knockout with lower activity and expression than wild-type aconitase. Transcriptome sequencing (RNA-seq) analysis of FA19 penA41 acnBG348D revealed a large set of upregulated genes involved in carbon and energy metabolism. We conclude that compensatory mutations can be selected in Cror gonococcal strains that increase metabolism to ameliorate their fitness deficit. IMPORTANCE The emergence of ceftriaxone-resistant (Cror) Neisseria gonorrhoeae has led to the looming threat of untreatable gonorrhea. Whether Cro resistance is likely to spread can be predicted from studies that compare the relative fitnesses of susceptible and resistant strains that differ only in the penA gene that confers Cro resistance. We showed that mosaic penA alleles found in Cror clinical isolates are outcompeted by the Cros parent strain in vitro and in vivo but that compensatory mutations that allow ceftriaxone resistance to be maintained by increasing bacterial fitness are selected during mouse infection. One compensatory mutant that was studied in more detail had a mutation in acnB, which encodes the aconitase that functions in the tricarboxylic acid (TCA) cycle. This study illustrates that compensatory mutations can be selected during infection, which we hypothesize may allow the spread of Cro resistance in nature. This study also provides novel insights into gonococcal metabolism and physiology.Leah R. VincentSamuel R. KerrYang TanJoshua TombergErica L. RatermanJulie C. Dunning HotoppMagnus UnemoRobert A. NicholasAnn E. JerseAmerican Society for MicrobiologyarticleNeisseria gonorrhoeaeaconitaseantibiotic resistancebiological fitnesscompetitive indexMicrobiologyQR1-502ENmBio, Vol 9, Iss 2 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Neisseria gonorrhoeae aconitase antibiotic resistance biological fitness competitive index Microbiology QR1-502
spellingShingle	Neisseria gonorrhoeae aconitase antibiotic resistance biological fitness competitive index Microbiology QR1-502 Leah R. Vincent Samuel R. Kerr Yang Tan Joshua Tomberg Erica L. Raterman Julie C. Dunning Hotopp Magnus Unemo Robert A. Nicholas Ann E. Jerse <italic toggle="yes">In Vivo</italic>-Selected Compensatory Mutations Restore the Fitness Cost of Mosaic <italic toggle="yes">penA</italic> Alleles That Confer Ceftriaxone Resistance in <italic toggle="yes">Neisseria gonorrhoeae</italic>
description	ABSTRACT Resistance to ceftriaxone in Neisseria gonorrhoeae is mainly conferred by mosaic penA alleles that encode penicillin-binding protein 2 (PBP2) variants with markedly lower rates of acylation by ceftriaxone. To assess the impact of these mosaic penA alleles on gonococcal fitness, we introduced the mosaic penA alleles from two ceftriaxone-resistant (Cror) clinical isolates (H041 and F89) into a Cros strain (FA19) by allelic exchange and showed that the resultant Cror mutants were significantly outcompeted by the Cros parent strain in vitro and in a murine infection model. Four Cror compensatory mutants of FA19 penA41 were isolated independently from mice that outcompeted the parent strain both in vitro and in vivo. One of these compensatory mutants (LV41C) displayed a unique growth profile, with rapid log growth followed by a sharp plateau/gradual decline at stationary phase. Genome sequencing of LV41C revealed a mutation (G348D) in the acnB gene encoding the bifunctional aconitate hydratase 2/2 methylisocitrate dehydratase. Introduction of the acnBG348D allele into FA19 penA41 conferred both a growth profile that phenocopied that of LV41C and a fitness advantage, although not as strongly as that exhibited by the original compensatory mutant, suggesting the existence of additional compensatory mutations. The mutant aconitase appears to be a functional knockout with lower activity and expression than wild-type aconitase. Transcriptome sequencing (RNA-seq) analysis of FA19 penA41 acnBG348D revealed a large set of upregulated genes involved in carbon and energy metabolism. We conclude that compensatory mutations can be selected in Cror gonococcal strains that increase metabolism to ameliorate their fitness deficit. IMPORTANCE The emergence of ceftriaxone-resistant (Cror) Neisseria gonorrhoeae has led to the looming threat of untreatable gonorrhea. Whether Cro resistance is likely to spread can be predicted from studies that compare the relative fitnesses of susceptible and resistant strains that differ only in the penA gene that confers Cro resistance. We showed that mosaic penA alleles found in Cror clinical isolates are outcompeted by the Cros parent strain in vitro and in vivo but that compensatory mutations that allow ceftriaxone resistance to be maintained by increasing bacterial fitness are selected during mouse infection. One compensatory mutant that was studied in more detail had a mutation in acnB, which encodes the aconitase that functions in the tricarboxylic acid (TCA) cycle. This study illustrates that compensatory mutations can be selected during infection, which we hypothesize may allow the spread of Cro resistance in nature. This study also provides novel insights into gonococcal metabolism and physiology.
format	article
author	Leah R. Vincent Samuel R. Kerr Yang Tan Joshua Tomberg Erica L. Raterman Julie C. Dunning Hotopp Magnus Unemo Robert A. Nicholas Ann E. Jerse
author_facet	Leah R. Vincent Samuel R. Kerr Yang Tan Joshua Tomberg Erica L. Raterman Julie C. Dunning Hotopp Magnus Unemo Robert A. Nicholas Ann E. Jerse
author_sort	Leah R. Vincent
title	<italic toggle="yes">In Vivo</italic>-Selected Compensatory Mutations Restore the Fitness Cost of Mosaic <italic toggle="yes">penA</italic> Alleles That Confer Ceftriaxone Resistance in <italic toggle="yes">Neisseria gonorrhoeae</italic>
title_short	<italic toggle="yes">In Vivo</italic>-Selected Compensatory Mutations Restore the Fitness Cost of Mosaic <italic toggle="yes">penA</italic> Alleles That Confer Ceftriaxone Resistance in <italic toggle="yes">Neisseria gonorrhoeae</italic>
title_full	<italic toggle="yes">In Vivo</italic>-Selected Compensatory Mutations Restore the Fitness Cost of Mosaic <italic toggle="yes">penA</italic> Alleles That Confer Ceftriaxone Resistance in <italic toggle="yes">Neisseria gonorrhoeae</italic>
title_fullStr	<italic toggle="yes">In Vivo</italic>-Selected Compensatory Mutations Restore the Fitness Cost of Mosaic <italic toggle="yes">penA</italic> Alleles That Confer Ceftriaxone Resistance in <italic toggle="yes">Neisseria gonorrhoeae</italic>
title_full_unstemmed	<italic toggle="yes">In Vivo</italic>-Selected Compensatory Mutations Restore the Fitness Cost of Mosaic <italic toggle="yes">penA</italic> Alleles That Confer Ceftriaxone Resistance in <italic toggle="yes">Neisseria gonorrhoeae</italic>
title_sort	<italic toggle="yes">in vivo</italic>-selected compensatory mutations restore the fitness cost of mosaic <italic toggle="yes">pena</italic> alleles that confer ceftriaxone resistance in <italic toggle="yes">neisseria gonorrhoeae</italic>
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/bf5cc6746a2948e9876c44e5b14525ce
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<italic toggle="yes">In Vivo</italic>-Selected Compensatory Mutations Restore the Fitness Cost of Mosaic <italic toggle="yes">penA</italic> Alleles That Confer Ceftriaxone Resistance in <italic toggle="yes">Neisseria gonorrhoeae</italic>

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