Upregulation of the phthiocerol dimycocerosate biosynthetic pathway by Rifampin-resistant, rpoB mutant Mycobacterium tuberculosis

Gregory P. Bisson, Carolina Mehaffy, Corey Broeckling, Jessica Prenni, Dalin Rifat, Desmond Lun, Marcos Burgos, Drew Weissman, C. Karakousis Petros, Karen Dobosc

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

Multidrug-resistant tuberculosis has emerged as a major threat to tuberculosis control. Phylogenetically related rifampin-resistant actinomycetes with mutations mapping to clinically dominant Mycobacterium tuberculosis mutations in the rpoB gene show upregulation of gene networks encoding secondary metabolites. We compared the expressed proteomes and metabolomes of two fully drug-susceptible clinical strains of M. tuberculosis (wild type) to those of their respective rifampin-resistant, rpoB mutant progeny strains with confirmed rifampin monoresistance following antitubercular therapy. Each of these strains was also used to infect gamma interferon- and lipopolysaccharide-activated murine J774A.1 macrophages to analyze transcriptional responses in a physiologically relevant model. Both rpoB mutants showed significant upregulation of the polyketide synthase genes ppsA-ppsE and drrA, which constitute an operon encoding multifunctional enzymes involved in the biosynthesis of phthiocerol dimycocerosate and other lipids in M. tuberculosis, but also of various secondary metabolites in related organisms, including antibiotics, such as erythromycin and rifamycins. ppsA (Rv2931), ppsB (Rv2932), and ppsC (Rv2933) were also found to be upregulated more than 10-fold in the Beijing rpoB mutant strain relative to its wild-type parent strain during infection of activated murine macrophages. In addition, metabolomics identified precursors of phthiocerol dimycocerosate, but not the intact molecule itself, in greater abundance in both rpoB mutant isolates. These data suggest that rpoB mutation in M. tuberculosis may trigger compensatory transcriptional changes in secondary metabolism genes analogous to those observed in related actinobacteria. These findings may assist in developing novel methods to diagnose and treat drug-resistant M. tuberculosis infections.

Original languageEnglish (US)
Pages (from-to)6441-6452
Number of pages12
JournalJournal of bacteriology
Volume194
Issue number23
DOIs
StatePublished - Dec 1 2012

Fingerprint

Biosynthetic Pathways
Rifampin
Mycobacterium tuberculosis
Up-Regulation
Multidrug-Resistant Tuberculosis
Actinobacteria
Mutation
Rifamycins
Macrophages
Multifunctional Enzymes
Polyketide Synthases
Secondary Metabolism
Genes
Mycobacterium Infections
Metabolomics
Metabolome
Gene Regulatory Networks
Erythromycin
Proteome
Operon

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology

Cite this

Bisson, Gregory P. ; Mehaffy, Carolina ; Broeckling, Corey ; Prenni, Jessica ; Rifat, Dalin ; Lun, Desmond ; Burgos, Marcos ; Weissman, Drew ; Petros, C. Karakousis ; Dobosc, Karen. / Upregulation of the phthiocerol dimycocerosate biosynthetic pathway by Rifampin-resistant, rpoB mutant Mycobacterium tuberculosis. In: Journal of bacteriology. 2012 ; Vol. 194, No. 23. pp. 6441-6452.
@article{76a857f608ba41bb9e85cae0c48ec1be,
title = "Upregulation of the phthiocerol dimycocerosate biosynthetic pathway by Rifampin-resistant, rpoB mutant Mycobacterium tuberculosis",
abstract = "Multidrug-resistant tuberculosis has emerged as a major threat to tuberculosis control. Phylogenetically related rifampin-resistant actinomycetes with mutations mapping to clinically dominant Mycobacterium tuberculosis mutations in the rpoB gene show upregulation of gene networks encoding secondary metabolites. We compared the expressed proteomes and metabolomes of two fully drug-susceptible clinical strains of M. tuberculosis (wild type) to those of their respective rifampin-resistant, rpoB mutant progeny strains with confirmed rifampin monoresistance following antitubercular therapy. Each of these strains was also used to infect gamma interferon- and lipopolysaccharide-activated murine J774A.1 macrophages to analyze transcriptional responses in a physiologically relevant model. Both rpoB mutants showed significant upregulation of the polyketide synthase genes ppsA-ppsE and drrA, which constitute an operon encoding multifunctional enzymes involved in the biosynthesis of phthiocerol dimycocerosate and other lipids in M. tuberculosis, but also of various secondary metabolites in related organisms, including antibiotics, such as erythromycin and rifamycins. ppsA (Rv2931), ppsB (Rv2932), and ppsC (Rv2933) were also found to be upregulated more than 10-fold in the Beijing rpoB mutant strain relative to its wild-type parent strain during infection of activated murine macrophages. In addition, metabolomics identified precursors of phthiocerol dimycocerosate, but not the intact molecule itself, in greater abundance in both rpoB mutant isolates. These data suggest that rpoB mutation in M. tuberculosis may trigger compensatory transcriptional changes in secondary metabolism genes analogous to those observed in related actinobacteria. These findings may assist in developing novel methods to diagnose and treat drug-resistant M. tuberculosis infections.",
author = "Bisson, {Gregory P.} and Carolina Mehaffy and Corey Broeckling and Jessica Prenni and Dalin Rifat and Desmond Lun and Marcos Burgos and Drew Weissman and Petros, {C. Karakousis} and Karen Dobosc",
year = "2012",
month = "12",
day = "1",
doi = "10.1128/JB.01013-12",
language = "English (US)",
volume = "194",
pages = "6441--6452",
journal = "Journal of Bacteriology",
issn = "0021-9193",
publisher = "American Society for Microbiology",
number = "23",

}

Bisson, GP, Mehaffy, C, Broeckling, C, Prenni, J, Rifat, D, Lun, D, Burgos, M, Weissman, D, Petros, CK & Dobosc, K 2012, 'Upregulation of the phthiocerol dimycocerosate biosynthetic pathway by Rifampin-resistant, rpoB mutant Mycobacterium tuberculosis', Journal of bacteriology, vol. 194, no. 23, pp. 6441-6452. https://doi.org/10.1128/JB.01013-12

Upregulation of the phthiocerol dimycocerosate biosynthetic pathway by Rifampin-resistant, rpoB mutant Mycobacterium tuberculosis. / Bisson, Gregory P.; Mehaffy, Carolina; Broeckling, Corey; Prenni, Jessica; Rifat, Dalin; Lun, Desmond; Burgos, Marcos; Weissman, Drew; Petros, C. Karakousis; Dobosc, Karen.

In: Journal of bacteriology, Vol. 194, No. 23, 01.12.2012, p. 6441-6452.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Upregulation of the phthiocerol dimycocerosate biosynthetic pathway by Rifampin-resistant, rpoB mutant Mycobacterium tuberculosis

AU - Bisson, Gregory P.

AU - Mehaffy, Carolina

AU - Broeckling, Corey

AU - Prenni, Jessica

AU - Rifat, Dalin

AU - Lun, Desmond

AU - Burgos, Marcos

AU - Weissman, Drew

AU - Petros, C. Karakousis

AU - Dobosc, Karen

PY - 2012/12/1

Y1 - 2012/12/1

N2 - Multidrug-resistant tuberculosis has emerged as a major threat to tuberculosis control. Phylogenetically related rifampin-resistant actinomycetes with mutations mapping to clinically dominant Mycobacterium tuberculosis mutations in the rpoB gene show upregulation of gene networks encoding secondary metabolites. We compared the expressed proteomes and metabolomes of two fully drug-susceptible clinical strains of M. tuberculosis (wild type) to those of their respective rifampin-resistant, rpoB mutant progeny strains with confirmed rifampin monoresistance following antitubercular therapy. Each of these strains was also used to infect gamma interferon- and lipopolysaccharide-activated murine J774A.1 macrophages to analyze transcriptional responses in a physiologically relevant model. Both rpoB mutants showed significant upregulation of the polyketide synthase genes ppsA-ppsE and drrA, which constitute an operon encoding multifunctional enzymes involved in the biosynthesis of phthiocerol dimycocerosate and other lipids in M. tuberculosis, but also of various secondary metabolites in related organisms, including antibiotics, such as erythromycin and rifamycins. ppsA (Rv2931), ppsB (Rv2932), and ppsC (Rv2933) were also found to be upregulated more than 10-fold in the Beijing rpoB mutant strain relative to its wild-type parent strain during infection of activated murine macrophages. In addition, metabolomics identified precursors of phthiocerol dimycocerosate, but not the intact molecule itself, in greater abundance in both rpoB mutant isolates. These data suggest that rpoB mutation in M. tuberculosis may trigger compensatory transcriptional changes in secondary metabolism genes analogous to those observed in related actinobacteria. These findings may assist in developing novel methods to diagnose and treat drug-resistant M. tuberculosis infections.

AB - Multidrug-resistant tuberculosis has emerged as a major threat to tuberculosis control. Phylogenetically related rifampin-resistant actinomycetes with mutations mapping to clinically dominant Mycobacterium tuberculosis mutations in the rpoB gene show upregulation of gene networks encoding secondary metabolites. We compared the expressed proteomes and metabolomes of two fully drug-susceptible clinical strains of M. tuberculosis (wild type) to those of their respective rifampin-resistant, rpoB mutant progeny strains with confirmed rifampin monoresistance following antitubercular therapy. Each of these strains was also used to infect gamma interferon- and lipopolysaccharide-activated murine J774A.1 macrophages to analyze transcriptional responses in a physiologically relevant model. Both rpoB mutants showed significant upregulation of the polyketide synthase genes ppsA-ppsE and drrA, which constitute an operon encoding multifunctional enzymes involved in the biosynthesis of phthiocerol dimycocerosate and other lipids in M. tuberculosis, but also of various secondary metabolites in related organisms, including antibiotics, such as erythromycin and rifamycins. ppsA (Rv2931), ppsB (Rv2932), and ppsC (Rv2933) were also found to be upregulated more than 10-fold in the Beijing rpoB mutant strain relative to its wild-type parent strain during infection of activated murine macrophages. In addition, metabolomics identified precursors of phthiocerol dimycocerosate, but not the intact molecule itself, in greater abundance in both rpoB mutant isolates. These data suggest that rpoB mutation in M. tuberculosis may trigger compensatory transcriptional changes in secondary metabolism genes analogous to those observed in related actinobacteria. These findings may assist in developing novel methods to diagnose and treat drug-resistant M. tuberculosis infections.

UR - http://www.scopus.com/inward/record.url?scp=84871057692&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84871057692&partnerID=8YFLogxK

U2 - 10.1128/JB.01013-12

DO - 10.1128/JB.01013-12

M3 - Article

VL - 194

SP - 6441

EP - 6452

JO - Journal of Bacteriology

JF - Journal of Bacteriology

SN - 0021-9193

IS - 23

ER -