Droplet Tn-Seq identifies the primary secretion mechanism for yersiniabactin in Yersinia pestis

Sarah L. Price, Derek Thibault, Taylor M. Garrison, Amanda Brady, Haixun Guo, Thomas E. Kehl-Fie, Sylvie Garneau-Tsodikova, Robert D. Perry, Tim van Opijnen, Matthew B. Lawrenz

Research output: Contribution to journalArticlepeer-review

Abstract

Nutritional immunity includes sequestration of transition metals from invading pathogens. Yersinia pestis overcomes nutritional immunity by secreting yersiniabactin to acquire iron and zinc during infection. While the mechanisms for yersiniabactin synthesis and import are well-defined, those responsible for yersiniabactin secretion are unknown. Identification of this mechanism has been difficult because conventional mutagenesis approaches are unable to inhibit trans-complementation by secreted factors between mutants. To overcome this obstacle, we utilized a technique called droplet Tn-seq (dTn-seq), which uses microfluidics to isolate individual transposon mutants in oil droplets, eliminating trans-complementation between bacteria. Using this approach, we first demonstrated the applicability of dTn-seq to identify genes with secreted functions. We then applied dTn-seq to identify an AcrAB efflux system as required for growth in metal-limited conditions. Finally, we showed this efflux system is the primary yersiniabactin secretion mechanism and required for virulence during bubonic and pneumonic plague. Together, these studies have revealed the yersiniabactin secretion mechanism that has eluded researchers for over 30 years and identified a potential therapeutic target for bacteria that use yersiniabactin for metal acquisition.

Original languageEnglish (US)
Article numbere57369
JournalEMBO Reports
Volume24
Issue number10
DOIs
StatePublished - Oct 9 2023

Keywords

  • Yersinia pestis
  • drug efflux systems
  • plague
  • siderophores
  • transposon mutagenesis

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology
  • Biochemistry

Fingerprint

Dive into the research topics of 'Droplet Tn-Seq identifies the primary secretion mechanism for yersiniabactin in Yersinia pestis'. Together they form a unique fingerprint.

Cite this