An RpoB mutation confers dual heteroresistance to daptomycin and vancomycin in Staphylococcus aureus

Longzhu Cui, Taisuke Isii, Minoru Fukuda, Tomonori Ochiai, Hui Min Neoh, Ilana Lopes Baratella Da Cunha Camargo, Yukiko Watanabe, Mitsutaka Shoji, Tomomi Hishinuma, Keiichi Hiramatsu

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Abstract

We have previously reported the establishment of a Staphylococcus aureus laboratory strain, 10*3d1, having reduced susceptibility to daptomycin and heterogeneous vancomycin-intermediate S. aureus (VISA) phenotype. The strain was generated in vitro by serial daptomycin selection (Camargo, I. L., H. M. Neoh, L. Cui, and K. Hiramatsu, Antimicrob. Agents Chemother. 52:4289-4299, 2008). Here we explored the genetic mechanism of resistance in the strain by whole-genome sequencing and by producing gene-replaced strains. By genome comparison between 10*3d1 and its parent methicillin-resistant Staphylococcus aureus (MRSA) strain N315ΔIP, we identified five nonsynonymous single nucleotide polymorphisms (SNPs). One of the five mutations was found in the rpoB gene encoding the RNA polymerase β subunit. The mutation at nucleotide position 1862 substituted the 621st alanine by glutamic acid. The replacement of the intact rpoB with the mutated rpoB, designated rpoB(A621E), conferred N315ΔIP with the phenotypes of reduced susceptibility to daptomycin and hetero-VISA. The rpoB(A621E)-mediated resistance conversion was accompanied by a thickened cell wall and reduction of the cell surface negative charge. Being consistent with these phenotypic changes, microarray data showed that the expression of the dlt operon, which increases the cell surface positive charge, was enhanced in the rpoB(A621E) mutant. Other remarkable findings of microarray analysis of the rpoB(A621E) mutant included repression of metabolic pathways of purine, pyrimidine, arginine, the urea cycle, and the lac operon, enhancement of the biosynthetic pathway of vitamin B2, K1, and K2, and cell wall metabolism. Finally, mutations identified in rplV and rplC, encoding 50S ribosomal proteins L22 and L3, respectively, were found to be associated with the slow growth, but not with the phenotype of decreased susceptibility to vancomycin and daptomycin, of 10*3d1.

Original languageEnglish
Pages (from-to)5222-5233
Number of pages12
JournalAntimicrobial Agents and Chemotherapy
Volume54
Issue number12
DOIs
Publication statusPublished - Dec 2010

Fingerprint

Daptomycin
Vancomycin
Staphylococcus aureus
Mutation
Phenotype
Cell Wall
Genome
Vitamin K 1
Vitamin K 2
Lac Operon
Riboflavin
Biosynthetic Pathways
DNA-Directed RNA Polymerases
Microarray Analysis
Operon
Methicillin-Resistant Staphylococcus aureus
Metabolic Networks and Pathways
Alanine
Genes
Single Nucleotide Polymorphism

ASJC Scopus subject areas

  • Pharmacology (medical)
  • Pharmacology
  • Infectious Diseases

Cite this

An RpoB mutation confers dual heteroresistance to daptomycin and vancomycin in Staphylococcus aureus. / Cui, Longzhu; Isii, Taisuke; Fukuda, Minoru; Ochiai, Tomonori; Neoh, Hui Min; Da Cunha Camargo, Ilana Lopes Baratella; Watanabe, Yukiko; Shoji, Mitsutaka; Hishinuma, Tomomi; Hiramatsu, Keiichi.

In: Antimicrobial Agents and Chemotherapy, Vol. 54, No. 12, 12.2010, p. 5222-5233.

Research output: Contribution to journalArticle

Cui, L, Isii, T, Fukuda, M, Ochiai, T, Neoh, HM, Da Cunha Camargo, ILB, Watanabe, Y, Shoji, M, Hishinuma, T & Hiramatsu, K 2010, 'An RpoB mutation confers dual heteroresistance to daptomycin and vancomycin in Staphylococcus aureus', Antimicrobial Agents and Chemotherapy, vol. 54, no. 12, pp. 5222-5233. https://doi.org/10.1128/AAC.00437-10
Cui, Longzhu ; Isii, Taisuke ; Fukuda, Minoru ; Ochiai, Tomonori ; Neoh, Hui Min ; Da Cunha Camargo, Ilana Lopes Baratella ; Watanabe, Yukiko ; Shoji, Mitsutaka ; Hishinuma, Tomomi ; Hiramatsu, Keiichi. / An RpoB mutation confers dual heteroresistance to daptomycin and vancomycin in Staphylococcus aureus. In: Antimicrobial Agents and Chemotherapy. 2010 ; Vol. 54, No. 12. pp. 5222-5233.
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