Skip to main content
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems
  • Log in
  • My alerts
  • My Cart

Main menu

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • Archive
    • Minireviews
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About AAC
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems

User menu

  • Log in
  • My alerts
  • My Cart

Search

  • Advanced search
Antimicrobial Agents and Chemotherapy
publisher-logosite-logo

Advanced Search

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • Archive
    • Minireviews
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About AAC
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
Mechanisms of Resistance

Pseudomonas aeruginosa High-Level Resistance to Polymyxins and Other Antimicrobial Peptides Requires cprA, a Gene That Is Disrupted in the PAO1 Strain

Alina D. Gutu, Nicole S. Rodgers, Jihye Park, Samuel M. Moskowitz
Alina D. Gutu
Department of Pediatrics, Massachusetts General Hospital, Boston, Massachusetts, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Nicole S. Rodgers
Department of Pediatrics, Massachusetts General Hospital, Boston, Massachusetts, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jihye Park
Department of Pediatrics, Massachusetts General Hospital, Boston, Massachusetts, USADepartment of Pediatrics, Harvard Medical School, Boston, Massachusetts, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Samuel M. Moskowitz
Department of Pediatrics, Massachusetts General Hospital, Boston, Massachusetts, USADepartment of Pediatrics, Harvard Medical School, Boston, Massachusetts, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1128/AAC.00904-15
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Tables
  • Additional Files
  • FIG 1
    • Open in new tab
    • Download powerpoint
    FIG 1

    Effect of phoPQ, pmrAB, or phoPQ and pmrAB deletion on Pm resistance in a PAK ΔcprRS cprRS21 background. For this and subsequent figures, Pm resistance experiments were performed twice; if discrepancies were seen, the experiment was performed a third time. Each panel shows a representative experiment, with the results expressed as means from three technical replicates. The error bars represent the standard deviations (SD). The nonitalicized allele name indicates the presence of an episomal version, i.e., an inducible expression strain (ctrl, empty vector control). Results from a PMB plate assay of strains induced with 0.1% l-Ara for 24 h are shown for strain 4537 (ΔphoPQ ΔcprRS cprRS21) with strain 4240 (ΔcprRS cprRS21) as a positive control and strain 4541 (ΔphoPQ ΔcprRS ctrl) as a negative control (A), strain 4472 (ΔpmrAB ΔcprRS cprRS21) with strain 4240 (ΔcprRS cprRS21) as a positive control and strain 4470 (ΔpmrAB ΔcprRS ctrl) as a negative control (B), and strain 4543 (ΔphoPQ ΔpmrAB ΔcprRS cprRS21) with strain 4240 (ΔcprRS cprRS21) as a positive control and strain 4547 (ΔphoPQ ΔpmrAB ΔcprRS ctrl) as a negative control (C).

  • FIG 2
    • Open in new tab
    • Download powerpoint
    FIG 2

    Effect of cprA2 deletion on Pm resistance in various PAK strain backgrounds. Shown are the results of a PMB plate assay of strain 4374 (ΔcprA2 ΔphoQ) and strain 4614 (ΔcprA2 ΔphoQ cprA2) induced with 0.1% l-Ara for 24 h, with strain 2326 (ΔphoQ) as a positive control (A), strain 4519 (ΔcprA2 ΔpmrAB pmrAB12) induced with 0.1% l-Ara for 24 h, with strain 2735 (ΔpmrAB pmrAB12) as a positive control (B), and strain 4425 (ΔcprA2 ΔcprRS cprRS21) induced with 0.1% l-Ara for 24 h, with strain 4240 (ΔcprRS cprRS21) as a positive control (C). Empty vector control strains 4497 (ΔcprA2 ΔphoQ pJN105), 4521 (ΔcprA2 ΔpmrAB pJN105), and 4447 (ΔcprA2 ΔcprRS pJN105) had Pm susceptibility values similar to those of the corresponding parental strains (not shown).

  • FIG 3
    • Open in new tab
    • Download powerpoint
    FIG 3

    Repair of the cprA1 mutant allele promotes high-level Pm resistance in the PAO1 ΔpmrAB strain expressing pmrAB12 in trans. Shown are the results of a PMB plate assay for strain 4763 (ΔcprA1::cprA+ ΔpmrAB pmrAB12) induced with 0.1% l-Ara for 24 h, with strains 4734 (ΔpmrAB pmrAB12) and 4765 (ΔcprA1::cprA+ ΔpmrAB ctrl) as negative controls.

  • FIG 4
    • Open in new tab
    • Download powerpoint
    FIG 4

    Deletion of cprA2 does not affect l-Ara4N modification of lipid A from the PAK strain. (A) Structure of lipid A with baseline hexa-acylation (m/z, 1,617), single l-Ara4N addition (m/z, 1,748), and double l-Ara4N addition (m/z, 1,879). Also shown are the MALDI-TOF spectra of lipid A isolated from strain 1026 (WT) (B), strain 2326 (ΔphoQ) (C), and strain 4374 (ΔcprA2 ΔphoQ) (D).

  • FIG 5
    • Open in new tab
    • Download powerpoint
    FIG 5

    Effect of arnC deletion in the PAK ΔcprRS cprRS21 strain. Shown are the results of a PMB plate assay of strain 4581 (ΔarnC ΔcprRS cprRS21) induced with 0.1% l-Ara for 24 h, with strain 4240 (ΔcprRS cprRS21) as a positive control and strain 4586 (ΔarnC ΔcprRS ctrl) as a negative control.

  • FIG 6
    • Open in new tab
    • Download powerpoint
    FIG 6

    Two-component regulatory systems mediating resistance to Pm and other CAPs modulate expression of arn and eptA, loci that enable specific lipid A modifications, and cprA, a locus potentially implicated in other LPS modification(s) and vesiculation from the outer membrane.

Tables

  • Figures
  • Additional Files
  • TABLE 1

    Strains of P. aeruginosa used in this work

    StrainStrain description (source/resistance profile)Reference
    1026PAK laboratory-adapted WT (S. Lory)65
    1027PAO1 laboratory-adapted WT (S. Lory)46
    1555PAO1 laboratory-adapted WT (M. Franklin)66
    2114PAO1 laboratory-adapted WT (C. Manoil)67
    2178PAO1 laboratory-adapted WT (M. Vasil)68
    1016Clinical isolate (Aarhus CF patient 2, 1996)12
    1020Clinical isolate (Copenhagen CF patient 6, 1998)12
    1603Clinical isolate (Copenhagen CF patient 14, 2003)12
    1611Clinical isolate (Copenhagen CF patient 15, 2001)12
    4240PAK ΔcprRS pJN105D::cprR+S21 (GENr)29
    4537PAK ΔphoPQ ΔcprRS pJN105D::cprR+S21(GENr)This study
    4541PAK ΔphoPQ ΔcprRS pJN105 (GENr)This study
    4472PAK ΔpmrAB ΔcprRS pJN105D::cprR+S21 (GENr)This study
    4470PAK ΔpmrAB ΔcprRS pJN105 (GENr)This study
    4543PAK ΔphoPQ ΔpmrAB ΔcprRS pJN105D::cprR+S21 (GENr)This study
    4547PAK ΔphoPQ ΔpmrAB ΔcprRS pJN105 (GENr)This study
    4232PAK ΔpmrAB ΔcprRS pJN105D::pmrAB12 (GENr)This study
    4234PAK ΔpmrAB ΔcprRS pJN105 (GENr)This study
    4343PAK ΔcprA2This study
    4458PAK ΔcprA2 pJN105 (GENr)This study
    4460PAK ΔcprA2 pJN105D::cprA2 (GENr)This study
    2326PAK ΔphoQ24
    4374PAK ΔcprA2 ΔphoQThis study
    4614PAK ΔcprA2 ΔphoQ pJN105D::cprA2 (GENr)This study
    4497PAK ΔcprA2 ΔphoQ pJN105 (GENr)This study
    3420PAK ΔpmrAB pJN105 (GENr)This study
    2735PAK ΔpmrAB pJN105D::pmrAB12 (GENr)12
    4519PAK ΔcprA2 ΔpmrAB pJN105D::pmrAB12 (GENr)This study
    4521PAK ΔcprA2 ΔpmrAB pJN105 (GENr)This study
    4425PAK ΔcprA2 ΔcprRS pJN105D::cprR+S21 (GENr)This study
    4447PAK ΔcprA2 ΔcprRS pJN105 (GENr)This study
    4200PAK ΔcprRS ΔphoQ pJN105D::cprR+S21 (GENr)12
    4608PAK ΔcprA2 ΔcprRS ΔphoQ pJN105D::cprR+S21 (GENr)This study
    4612PAK ΔcprA2 ΔcprRS ΔphoQ pJN105 (GENr)This study
    4581PAK ΔarnC ΔcprRS pJN105D::cprR+S21(GENr)This study
    4586PAK ΔarnC ΔcprRS pJN105 (GENr)This study
    4734PAO1 ΔpmrAB pJN105D::pmrAB12 (GENr)This study
    4763PAO1 ΔpmrAB ΔcprA1::cprA+ pJN105D::pmrAB12 (GENr)This study
    4765PAO1 ΔpmrAB ΔcprA1::cprA+ pJN105 (GENr)This study
  • TABLE 2

    Effect of cprA2 deletion in the PAK strain on pmrAB12-induced cationic antimicrobial peptide resistance

    Strain/genotypePMBaProtegrin-1C18G
    EC50 (mg/liter)FIREC50 (mg/liter)FIREC50 (mg/liter)FIR
    PAK WT<0.12510.125121
    PAK ΔpmrAB pJN105<0.12510.125121
    PAK ΔpmrAB pJN105D::pmrAB1232>2561296256128
    PAK ΔcprA2 ΔpmrAB pJN105D::pmrAB122>162166432
    • ↵a EC50, effective concentration that kills 50% of CFU; FIR, fold increase in resistance to the indicated antimicrobial peptide of each strain with respect to PAK WT (ratio of EC50s).

  • TABLE 3

    Relative abundance of cprA and arnC mRNA transcripts in phoQ deletion and pmrAB12 or cprRS21 expression backgrounds, determined by RT-qPCR analysis (n = 3 biological replicates)

    GenotypeRelative mRNA abundance ± SDa
    cprAarnC
    PAK ΔphoQ1 ± 0.434 ± 17
    PAK ΔpmrAB pJN105D::pmrAB1294 ± 2621 ± 7
    PAK ΔcprRS pJN105D::cpRS21165 ± 3734 ± 18
    PAK ΔcprA2 ΔcprRS pJN105D::cpRS21NA30 ± 9
    PAK ΔarnC ΔcprRS pJN105D::cpRS21172 ± 70NA
    PAK ΔphoQ ΔcprRS pJN105D::cprRS21236 ± 16251 ± 40
    • ↵a Normalized to mRNA abundance in the PAK WT strain. NA, not applicable.

Additional Files

  • Figures
  • Tables
  • Supplemental material

    • Supplemental file 1 -

      Supplemental Tables S1 and S2 and Figures S1 to S4.

      PDF, 386K

PreviousNext
Back to top
Download PDF
Citation Tools
Pseudomonas aeruginosa High-Level Resistance to Polymyxins and Other Antimicrobial Peptides Requires cprA, a Gene That Is Disrupted in the PAO1 Strain
Alina D. Gutu, Nicole S. Rodgers, Jihye Park, Samuel M. Moskowitz
Antimicrobial Agents and Chemotherapy Aug 2015, 59 (9) 5377-5387; DOI: 10.1128/AAC.00904-15

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Print

Alerts
Sign In to Email Alerts with your Email Address
Email

Thank you for sharing this Antimicrobial Agents and Chemotherapy article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Pseudomonas aeruginosa High-Level Resistance to Polymyxins and Other Antimicrobial Peptides Requires cprA, a Gene That Is Disrupted in the PAO1 Strain
(Your Name) has forwarded a page to you from Antimicrobial Agents and Chemotherapy
(Your Name) thought you would be interested in this article in Antimicrobial Agents and Chemotherapy.
Share
Pseudomonas aeruginosa High-Level Resistance to Polymyxins and Other Antimicrobial Peptides Requires cprA, a Gene That Is Disrupted in the PAO1 Strain
Alina D. Gutu, Nicole S. Rodgers, Jihye Park, Samuel M. Moskowitz
Antimicrobial Agents and Chemotherapy Aug 2015, 59 (9) 5377-5387; DOI: 10.1128/AAC.00904-15
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Top
  • Article
    • ABSTRACT
    • INTRODUCTION
    • MATERIALS AND METHODS
    • RESULTS
    • DISCUSSION
    • ACKNOWLEDGMENTS
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

Related Articles

Cited By...

About

  • About AAC
  • Editor in Chief
  • Editorial Board
  • Policies
  • For Reviewers
  • For the Media
  • For Librarians
  • For Advertisers
  • Alerts
  • RSS
  • FAQ
  • Permissions
  • Journal Announcements

Authors

  • ASM Author Center
  • Submit a Manuscript
  • Article Types
  • Ethics
  • Contact Us

Follow #AACJournal

@ASMicrobiology

       

ASM Journals

ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology.

About ASM | Contact Us | Press Room

 

ASM is a member of

Scientific Society Publisher Alliance

Copyright © 2019 American Society for Microbiology | Privacy Policy | Website feedback

Print ISSN: 0066-4804; Online ISSN: 1098-6596