Modulation of rob expression accelerates development of antibiotic resistance in Yersinia enterocolitica

Abstract

Multidrug-resistant bacteria pose a severe threat to global health. Mutations in transcriptional regulators accelerate the emergence of multidrug resistance and may have a crucial impact on pathogen evolvability under antibiotic exposure. Here, we investigate these dynamics in the bacterial pathogen Yersinia enterocolitica. In this organism, we identified a high-frequency de novo mutation in the promoter of an AraC/XylS-family transcriptional regulator, Rob. This mutation arose independently during resistance evolution against three of six antibiotic classes. Sequence and structure alignments indicate that Rob is a previously uncharacterized, lineage-specific regulator in Y. enterocolitica, featuring a conserved promoter architecture. This promoter mutation resulted in robust rob overexpression, leading to the activation of multiple downstream efflux- and membrane-associated pathways. This regulatory mutation emerges early and shapes the acquisition of tetracycline resistance, while also enhancing high-level enrofloxacin resistance in combination with canonical mutations in gyrA and parC. Despite being favored under antibiotic selection, rob overexpression is costly, resulting in counter-selection of the -57 G>A rob allele in the absence of antibiotics. Together, these findings identify Rob as a Yersinia-specific efflux regulator and demonstrate how regulatory mutations can transiently accelerate antibiotic resistance. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=80 SRC="FIGDIR/small/707304v1_ufig1.gif" ALT="Figure 1"> View larger version (30K): org.highwire.dtl.DTLVardef@107aa88org.highwire.dtl.DTLVardef@4ccd66org.highwire.dtl.DTLVardef@411e2aorg.highwire.dtl.DTLVardef@1235fbc_HPS_FORMAT_FIGEXP M_FIG C_FIG