摘要：SummaryTrimethoprim, a preferred treatment for urinary tract infections, is becoming obsolete owing to the rapid dissemination of resistantE. coli. Although direct resistance mechanisms such as overexpression of a mutant FolA anddfrenzymes are well characterized, associated alterations that drive or sustain resistance are unknown. We identify the repertoire of resistance-associated perturbations by constructing and interrogating a transcriptome-integrated functional interactome. From the cross talk between perturbations in stress-response and metabolic pathways, we identify the critical dependence on serine hydroxymethyltransferase (GlyA) as an emergent vulnerability. Through its deletion, we demonstrate that GlyA is necessary to sustain high levels of resistance in both laboratory-evolved resistantE. coliand a multidrug-resistant clinical isolate. Through comparative evolution, we show that the absence of GlyA activity decelerates the acquisition of resistance inE. coli. Put together, our results identify GlyA as a promising target, providing a basis for the rational design of drug combinations.Graphical AbstractDisplay OmittedHighlights•TMP-resistantE. colishow cross talk between stress response and metabolic pathways•Dependence onglyAis an emergent vulnerability associated with TMP resistance•Knockout ofglyApartially rescues sensitivity to TMP inE. coliMicrobiology; Multi-Drug Resistant Organisms; Transcriptomics
