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
    • Latest Articles
    • COVID-19 Special Collection
    • Archive
    • Minireviews
  • Topics
    • Applied and Environmental Science
    • Clinical Science and Epidemiology
    • Ecological and Evolutionary Science
    • Host-Microbe Biology
    • Molecular Biology and Physiology
    • Therapeutics and Prevention
  • 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 mBio
    • Editor in Chief
    • Board of Editors
    • AAM Fellows
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • 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
mBio
publisher-logosite-logo

Advanced Search

  • Home
  • Articles
    • Latest Articles
    • COVID-19 Special Collection
    • Archive
    • Minireviews
  • Topics
    • Applied and Environmental Science
    • Clinical Science and Epidemiology
    • Ecological and Evolutionary Science
    • Host-Microbe Biology
    • Molecular Biology and Physiology
    • Therapeutics and Prevention
  • 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 mBio
    • Editor in Chief
    • Board of Editors
    • AAM Fellows
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
Research Article

The BvgAS Regulon of Bordetella pertussis

Kyung Moon, Richard P. Bonocora, David D. Kim, Qing Chen, Joseph T. Wade, Scott Stibitz, Deborah M. Hinton
Rino Rappuoli, Editor
Kyung Moon
a Gene Expression and Regulation Section, Laboratory of Cell and Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Richard P. Bonocora
b Wadsworth Center, New York State Department of Health, Albany, New York, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
David D. Kim
a Gene Expression and Regulation Section, Laboratory of Cell and Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Qing Chen
c Division of Bacterial, Parasitic and Allergenic Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, Maryland, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Joseph T. Wade
b Wadsworth Center, New York State Department of Health, Albany, New York, USA
d Department of Biomedical Sciences, School of Public Health, University at Albany, Albany, New York, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Scott Stibitz
c Division of Bacterial, Parasitic and Allergenic Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, Maryland, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Deborah M. Hinton
a Gene Expression and Regulation Section, Laboratory of Cell and Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Rino Rappuoli
GSK Vaccines
Roles: Editor
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
James Slauch
University of Illinois at Urbana Champaign
Roles: Solicited external reviewer
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Erik Hewlett
University of Virginia School of Medicine
Roles: Solicited external reviewer
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1128/mBio.01526-17
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Tables
  • Supplemental Material
  • FIG 1 
    • Open in new tab
    • Download powerpoint
    FIG 1 

    Relative expression of selected genes of transcriptional regulators positively regulated by BvgA~P. Values are shown as fold difference using the 2−ΔΔCT method (65). Error bars indicate standard deviations among three independent biological replicates for each condition with more than two different trials. Blue bars, FD of WT without MgSO4 versus WT with MgSO4; red bars, FD of WT without MgSO4 versus ΔbvgAS strain without MgSO4. Student’s t test was used for statistical analysis, and genes showing significant differences between WT without MgSO4 and WT with MgSO4 or between WT without MgSO4 and ΔbvgAS strain without MgSO4 are labeled as *** (P < 0.001), ** (P < 0.01), or * (P < 0.05). “ns” represents statistically nonsignificant.

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

    Relative expression of selected genes involved in metabolic pathways negatively affected by BvgA~P, as measured by RT-qPCR. Values are shown as fold difference using the 2−ΔΔCT method (65). Error bars indicate standard deviations among three independent biological replicates for each condition with more than two different trials. Blue bars, FD of WT without MgSO4 versus WT with MgSO4; red bars, FD of WT without MgSO4 versus ΔbvgAS strain without MgSO4. Bracketed genes are located in the same locus of the genome. Student’s t test was used for statistical analysis, and genes showing significant differences between WT without MgSO4 and WT with MgSO4 or between WT without MgSO4 and ΔbvgAS strain without MgSO4 are labeled as *** (P < 0.001), ** (P < 0.01), or * (P < 0.05).

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

    PbrpL is directly activated by BvgA~P. (A) Denaturing polyacrylamide gels showing RNA obtained after in vitro transcription of the indicated templates using (as indicated) B. pertussis RNAP alone (−), RNAP plus BvgA (+), or RNAP plus BvgA~P (P). The expected transcripts (nucleotides) are indicated. (B) Primer extension analysis of in vitro RNA to identify the start site of PbrpL. A denaturing gel of primer extension products is shown. Left lanes are sequencing ladders; right lanes show primer extension products for RNA obtained in the absence of BvgA (−), the presence of BvgA~P (P), or the presence of nonphosphorylated BvgA (+). (C) Sequence of the brpL promoter, determined from primer extension analyses in panel A, and of the promoters for BP1005 and BP3441, determined from transcript length in in vitro transcription in panel B. Assigned −10 and −35 elements (BP1005 and BP3441) are in blue; +1 is in red.

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

    FeBABE cleavage sites for PbrpL and PfhaB. (A) Denaturing gels showing products of FeBABE cleavage using complexes formed with PbrpL (left) or PfhaB (right), 32P end labeled on either the template or the nontemplate strand, B. pertussis RNAP, and (as indicated) either phosphorylated BvgAT194C-FeBABE or phosphorylated BvgAV148C-FeBABE. The G+A lanes correspond to DNA ladders generated by Maxam-Gilbert sequencing with the positions relative to the transcription start sites indicated (36). Arrowheads denote sites of cleavage. (B) Sequences of PbrpL and PfhaB with the cut sites generated by BvgAT194C-FeBABE (pink arrowheads) and by BvgAV148C-FeBABE (purple arrowheads). BvgA binding sites are boxed in orange; the −10 regions are shown and boxed in blue. (An alternative −10 element for PfhaB is indicated in the dashed purple box.) Transcription start sites (+1) are in red. Numbers above the inverted red arrows indicate the predicted score for BvgA binding half-sites in each promoter according to the algorithm reported by Merkel et al. (67). For instance, the sequence of the perfect inverted repeat is TAGGAAATTTCCTA, whose score is given as “0.”

Tables

  • Figures
  • Supplemental Material
  • TABLE 1 

    Genes of transcriptional regulators positively regulated by BvgA~Pa

    TABLE 1 
    • ↵a Expression of genes in boldface was checked by RT-qPCR. Underlined genes are newly identified in this study.

  • TABLE 2 

    RNA-seq expression levels of selected well-known vag genes and vrg genesa

    TABLE 2 
    • ↵a Expression of genes in bold was checked by RT-qPCR.

    • b FD represents the fold difference in expression of WT without MgSO4 versus ΔbvgAS strain without MgSO4.

Supplemental Material

  • Figures
  • Tables
  • TABLE S1 

    (A) Entire set of RNA-seq data. (B) Genes positively regulated by BvgA~P. Genes whose expressions have an FD of ≥1.6 with a P value of <0.05 are considered to be significant. Genes in blue are newly identified in this study; the gene in red is not Bvg(+) as indicated by RT-qPCR. Genes highlighted in yellow were assessed by RT-qPCR. 1Possible function identified by PSI-BLAST in this study. 2Possible function assigned in the work of Ahuja et al. (32). A total of 245 genes are identified as Bvg(+), 237 by RNA-seq and 8 from RT-qPCR. One hundred twenty-one genes are newly identified as Bvg(+) in this study. (C) Genes negatively regulated by BvgA~P. Genes whose expressions have an FD of ≥1.6 with a P value of <0.05 are considered to be significant. Genes in blue are newly identified in this study; the gene in red is not Bvg(−) as indicated by RT-qPCR. Genes highlighted in yellow were assessed by RT-qPCR. 1Possible function identified by PSI-BLAST in this study. A total of 326 genes are identified as Bvg(−), 321 by RNA-seq and 5 from RT-qPCR. Two hundred thirty-two genes are newly identified as Bvg(−) in this study. Download TABLE S1, XLSX file, 6.6 MB.

    This is a work of the U.S. Government and is not subject to copyright protection in the United States. Foreign copyrights may apply.

  • FIG S1 

    RT-qPCR analyses of genes identified as positively regulated by BvgA~P by the RNA-seq analyses. (A) Well-known vags; (B) T3SS genes; (C) genes of membrane-associated transporter proteins and hypothetical proteins. Values are shown as a fold difference using the 2−ΔΔCT method. Error bars indicate standard deviations among three independent biological replicates for each condition with more than two different trials. Blue bar, FD of WT without MgSO4 versus WT with MgSO4; red bar, FD of WT without MgSO4 versus ΔbvgAS without MgSO4. Download FIG S1, EPS file, 4.7 MB.

    This is a work of the U.S. Government and is not subject to copyright protection in the United States. Foreign copyrights may apply.

  • FIG S2 

    RT-qPCR analyses of genes identified as negatively regulated by BvgA~P by the RNA-seq analyses. (A) Well-known vrgs; (B) other vrgs, including genes for cold shock membrane proteins and genes with hypothetical functions. (Cold shock genes are bracketed.) Values are shown as a fold difference using the 2−ΔΔCT method. Error bars indicate standard deviations among three independent biological replicates for each condition with more than two different trials. Blue bar, FD of WT without MgSO4 versus WT with MgSO4; red bar, FD of WT without MgSO4 versus ΔbvgAS without MgSO4. Download FIG S2, EPS file, 3.1 MB.

    This is a work of the U.S. Government and is not subject to copyright protection in the United States. Foreign copyrights may apply.

  • TABLE S2 

    Well-known vags, T3SS genes, and genes of membrane-associated transporter proteins and hypothetical proteins. (A) Well-known vags positively regulated by BvgA~P; (B) T3SS genes positively regulated by BvgA~P; (C) genes of membrane-associated transporter proteins and hypothetical proteins positively regulated by BvgA~P. Download TABLE S2, DOCX file, 0.1 MB.

    This is a work of the U.S. Government and is not subject to copyright protection in the United States. Foreign copyrights may apply.

  • TABLE S3 

    Well-known vrgs and other vrgs, including genes for cold shock and membrane proteins and genes with hypothetical functions. (A) Well-known vrgs; (B) other vrgs, including genes for cold shock and membrane proteins and genes with hypothetical functions. Download TABLE S3, DOCX file, 0.03 MB.

    This is a work of the U.S. Government and is not subject to copyright protection in the United States. Foreign copyrights may apply.

  • TABLE S4 

    Genes involved in various metabolic pathways negatively regulated by BvgA~P. Download TABLE S4, DOCX file, 0.05 MB.

    This is a work of the U.S. Government and is not subject to copyright protection in the United States. Foreign copyrights may apply.

  • TABLE S5 

    RT-qPCR analysis for other possible vags and vrgs. Genes were previously identified as vags in the work of Cummings et al. (9) in B. pertussis Tohama I,1 in B. pertussis GMT-1,2 in B. bronchiseptica RB50,3 or in B. bronchiseptica 77.4,5 Genes were previously identified as vrgs in the work of Cummings et al. (9). (A) Possible vag transcriptional regulators, not identified by our RNA-seq; (B) genes reported as vrgs in B. bronchiseptica. Download TABLE S5, DOCX file, 0.02 MB.

    This is a work of the U.S. Government and is not subject to copyright protection in the United States. Foreign copyrights may apply.

  • FIG S3 

    Cleavage of PbrpL by FeBABE-conjugated BvgA~P in transcription complexes made with E. coli RNAP. (A) Portion of denaturing polyacrylamide gels showing PbrpL RNA (left) or PfhaB RNA (right) obtained after in vitro transcription using, as indicated, E. coli RNAP alone (-), RNAP plus BvgA (+), or RNAP plus BvgA~P (P). (B) Denaturing gels showing products of FeBABE cleavage using complexes formed with PbrpL (left) or PfhaB (right), 32P end labeled on either the template or the nontemplate strand, E. coli RNA polymerase, and, as indicated, either phosphorylated BvgAT194C-FeBABE or phosphorylated BvgAV148C-FeBABE. The G+A lanes correspond to DNA ladders generated by Maxam-Gilbert sequencing with the positions relative to the transcription start sites indicated. Arrowheads denote sites of cleavage. (C) Sequences of PbrpL and PfhaB with the cut sites generated by BvgAT194C-FeBABE (pink arrowheads) and by BvgAV148C-FeBABE (purple arrowheads). BvgA binding sites are boxed in orange; the −10 regions are boxed in blue. (An alternative −10 element for PfhaB is indicated in the dashed purple box.) Transcription start sites (+1) are in red. Numbers above the inverted red arrows indicate the predicted score for BvgA binding half-sites in each promoter according to the algorithm reported by Merkel et al. (67). Download FIG S3, TIF file, 25.1 MB.

    This is a work of the U.S. Government and is not subject to copyright protection in the United States. Foreign copyrights may apply.

PreviousNext
Back to top
Download PDF
Citation Tools
The BvgAS Regulon of Bordetella pertussis
Kyung Moon, Richard P. Bonocora, David D. Kim, Qing Chen, Joseph T. Wade, Scott Stibitz, Deborah M. Hinton
mBio Oct 2017, 8 (5) e01526-17; DOI: 10.1128/mBio.01526-17

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 mBio 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.
The BvgAS Regulon of Bordetella pertussis
(Your Name) has forwarded a page to you from mBio
(Your Name) thought you would be interested in this article in mBio.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
The BvgAS Regulon of Bordetella pertussis
Kyung Moon, Richard P. Bonocora, David D. Kim, Qing Chen, Joseph T. Wade, Scott Stibitz, Deborah M. Hinton
mBio Oct 2017, 8 (5) e01526-17; DOI: 10.1128/mBio.01526-17
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Top
  • Article
    • ABSTRACT
    • INTRODUCTION
    • RESULTS
    • DISCUSSION
    • MATERIALS AND METHODS
    • ACKNOWLEDGMENTS
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

KEYWORDS

Bacterial Proteins
Bordetella pertussis
Regulon
transcription factors
BvgAS regulon
RNA polymerase
RNA-seq
brpL
pertussis

Related Articles

Cited By...

About

  • About mBio
  • Editor in Chief
  • Board of Editors
  • AAM Fellows
  • Policies
  • For Reviewers
  • For the Media
  • For Librarians
  • For Advertisers
  • Alerts
  • RSS
  • FAQ
  • Permissions
  • Journal Announcements

Authors

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

Follow #mBio

@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

 

American Society for Microbiology
1752 N St. NW
Washington, DC 20036
Phone: (202) 737-3600

Copyright © 2021 American Society for Microbiology | Privacy Policy | Website feedback

Online ISSN: 2150-7511