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Mechanisms of Resistance

Mutation of RNA Polymerase β-Subunit Gene Promotes Heterogeneous-to-Homogeneous Conversion of β-Lactam Resistance in Methicillin-Resistant Staphylococcus aureus

Yoshifumi Aiba, Yuki Katayama, Tomomi Hishinuma, Hiroko Murakami-Kuroda, Longzhu Cui, Keiichi Hiramatsu
Yoshifumi Aiba
Department of Infection Control Science, Graduate School of Medicine, Juntendo University, Tokyo, Japana
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Yuki Katayama
Department of Bacteriology, Faculty of Medicine, Juntendo University,b Tokyo, Japan
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Tomomi Hishinuma
Department of Bacteriology, Faculty of Medicine, Juntendo University,b Tokyo, Japan
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Hiroko Murakami-Kuroda
Department of Bacteriology, Faculty of Medicine, Juntendo University,b Tokyo, Japan
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Longzhu Cui
Department of Bacteriology, Faculty of Medicine, Juntendo University,b Tokyo, Japan
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Keiichi Hiramatsu
Department of Infection Control Science, Graduate School of Medicine, Juntendo University, Tokyo, Japana
Department of Bacteriology, Faculty of Medicine, Juntendo University,b Tokyo, Japan
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DOI: 10.1128/AAC.00720-13
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ABSTRACT

Three types of phenotypic expression of β-lactam resistance have been reported in methicillin-resistant Staphylococcus aureus (MRSA): heterogeneous, homogeneous, and Eagle-type resistance. Heterogeneous-to-homogeneous conversion of β-lactam resistance is postulated to be caused by a chromosomal mutation (chr*) in addition to the expression of the mecA gene. Eagle-type resistance is a unique phenotype of chr* occurring in pre-MRSA strain N315 whose mecA gene expression is strongly repressed by an intact mecI gene. We here report that certain mutations of the rpoB gene, encoding the RNA polymerase β subunit, belong to chr*. We studied homogeneous MRSA (homo-MRSA) strain N315ΔIP-H5 (abbreviated as ΔIP-H5), which was obtained from hetero-MRSA strain N315ΔIP by selection with 8 mg/liter imipenem. Whole-genome sequencing of ΔIP-H5 revealed the presence of a unique mutation in the rpoB gene, rpoB(N967I), causing the amino acid replacement of Asn by Ile at position 967 of RpoB. The effect of the rpoB(N967I) mutation was confirmed by constructing a revertant H5 rpoB(I967N) strain as well as an N315-derived mutant, N315 rpoB(N967I). H5 rpoB(I967N) regained the hetero-resistance phenotype, and the N315 rpoB(N967I) strain showed an Eagle-type phenotype similar to that of the typical Eagle-type MRSA strain N315h4. Furthermore, subsequent whole-genome sequencing revealed that N315h4 also had a missense mutation of rpoB(R644H). Introduction of the rpoB(N967I) mutation was accompanied by decreased autolysis, prolonged doubling time, and tolerance to bactericidal concentrations of methicillin. We consider that rpoB mutations are the major cause for heterogeneous-to-homogeneous phenotypic conversion of β-lactam resistance in MRSA strain N315 and its derived strains.

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Mutation of RNA Polymerase β-Subunit Gene Promotes Heterogeneous-to-Homogeneous Conversion of β-Lactam Resistance in Methicillin-Resistant Staphylococcus aureus
Yoshifumi Aiba, Yuki Katayama, Tomomi Hishinuma, Hiroko Murakami-Kuroda, Longzhu Cui, Keiichi Hiramatsu
Antimicrobial Agents and Chemotherapy Sep 2013, 57 (10) 4861-4871; DOI: 10.1128/AAC.00720-13

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Mutation of RNA Polymerase β-Subunit Gene Promotes Heterogeneous-to-Homogeneous Conversion of β-Lactam Resistance in Methicillin-Resistant Staphylococcus aureus
Yoshifumi Aiba, Yuki Katayama, Tomomi Hishinuma, Hiroko Murakami-Kuroda, Longzhu Cui, Keiichi Hiramatsu
Antimicrobial Agents and Chemotherapy Sep 2013, 57 (10) 4861-4871; DOI: 10.1128/AAC.00720-13
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