Azithromycin resistance mutations in Streptococcus pneumoniae as revealed by a chemogenomic screen.
We report on the combination of chemical mutagenesis, azithromycin selection and next-generation sequencing (Mut-Seq) for the identification of small nucleotide variants that decrease the susceptibility of Streptococcus pneumoniae to the macrolide antibiotic azithromycin. Mutations in the 23S ribosomal RNA or in ribosomal proteins can confer resistance to macrolides and these were detected by Mut-Seq. By concentrating on recurrent variants, we could associate mutations in genes implicated in the metabolism of glutamine with decreased azithromycin susceptibility among S. pneumoniae mutants. Glutamine synthetase catalyses the transformation of glutamate and ammonium into glutamine and its chemical inhibition is shown to sensitize S. pneumoniae to antibiotics. A mutation affecting the ribosomal-binding site of a putative ribonuclease J2 is also shown to confer low-level resistance. Mut-Seq has the potential to reveal chromosomal changes enabling high resistance as well as novel events conferring more subtle phenotypes.
['Gingras|Hélène|H|', 'Patron|Kévin|K|', 'Leprohon|Philippe|P|', 'Ouellette|Marc|M|']
D000900:Anti-Bacterial Agents D017963:Azithromycin D024881:Drug Resistance, Bacterial D004722:Endoribonucleases D005974:Glutamate-Ammonia Ligase D059014:High-Throughput Nucleotide Sequencing D008826:Microbial Sensitivity Tests D016296:Mutagenesis D011500:Protein Synthesis Inhibitors D012338:RNA, Ribosomal, 23S D013296:Streptococcus pneumoniae
Azithromycin Resistance Streptococcus pneumoniae chemical mutagenesis macrolide Glutamine next generation sequencing