Evolutionary paths to macrolide resistance in a Neisseria commensal converge on ribosomal genes through short sequence duplications

PLoS One. 2022 Jan 13;17(1):e0262370. doi: 10.1371/journal.pone.0262370. eCollection 2022.

Abstract

Neisseria commensals are an indisputable source of resistance for their pathogenic relatives. However, the evolutionary paths commensal species take to reduced susceptibility in this genus have been relatively underexplored. Here, we leverage in vitro selection as a powerful screen to identify the genetic adaptations that produce azithromycin resistance (≥ 2 μg/mL) in the Neisseria commensal, N. elongata. Across multiple lineages (n = 7/16), we find mutations that reduce susceptibility to azithromycin converge on the locus encoding the 50S ribosomal L34 protein (rpmH) and the intergenic region proximal to the 30S ribosomal S3 protein (rpsC) through short tandem duplication events. Interestingly, one of the laboratory evolved mutations in rpmH is identical (7LKRTYQ12), and two nearly identical, to those recently reported to contribute to high-level azithromycin resistance in N. gonorrhoeae. Transformations into the ancestral N. elongata lineage confirmed the causality of both rpmH and rpsC mutations. Though most lineages inheriting duplications suffered in vitro fitness costs, one variant showed no growth defect, suggesting the possibility that it may be sustained in natural populations. Ultimately, studies like this will be critical for predicting commensal alleles that could rapidly disseminate into pathogen populations via allelic exchange across recombinogenic microbial genera.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • Azithromycin / pharmacology
  • Drug Resistance, Bacterial / genetics*
  • Macrolides / pharmacology*
  • Microbial Sensitivity Tests
  • Microbiota / genetics
  • Neisseria / genetics*
  • Protein Synthesis Inhibitors
  • RNA, Ribosomal, 23S / genetics
  • Ribosomal Proteins / genetics
  • Ribosome Subunits, Large, Bacterial / genetics
  • Ribosome Subunits, Small, Bacterial / genetics
  • Ribosomes / genetics
  • Sequence Deletion / genetics

Substances

  • Anti-Bacterial Agents
  • Macrolides
  • Protein Synthesis Inhibitors
  • RNA, Ribosomal, 23S
  • Ribosomal Proteins
  • ribosomal protein L34
  • ribosomal protein S3
  • ribosomal protein S30
  • Azithromycin

Grants and funding

The authors would like to acknowledge the generous support provided by the RIT College of Science for this study. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.