Functional metagenomic approach to identify overlooked antibiotic resistance mutations in bacterial rRNA

Kentaro Miyazaki, Kei Kitahara

Research output: Contribution to journalArticlepeer-review

4 Citations (Scopus)

Abstract

Our knowledge as to how bacteria acquire antibiotic resistance is still fragmented, especially for the ribosome-Targeting drugs. In this study, with the aim of finding novel mechanisms that render bacteria resistant to the ribosome-Targeting antibiotics, we developed a general method to systematically screen for antibiotic resistant 16 S ribosomal RNAs (rRNAs), which are the major target for multiple antibiotics (e.g. spectinomycin, tetracycline, and aminoglycosides), and identify point mutations therein. We used Escherichia coli 7, a null mutant of the rrn (ribosomal RNA) operons, as a surrogate host organism to construct a metagenomic library of 16 S rRNA genes from the natural (non-clinical) environment. The library was screened for spectinomycin resistance to obtain four resistant 16 S rRNA genes from non-E. coli bacterial species. Bioinformatic analysis and site-directed mutagenesis identified three novel mutations-U1183C (the first mutation discovered in a region other than helix 34), and C1063U and U1189C in helix 34-As well as three well-described mutations (C1066U, C1192G, and G1193A). These results strongly suggest that uncharacterized antibiotic resistance mutations still exist, even for traditional antibiotics.

Original languageEnglish
Article number5179
JournalScientific reports
Volume8
Issue number1
DOIs
Publication statusPublished - Dec 1 2018
Externally publishedYes

ASJC Scopus subject areas

  • General

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