Exploring antibiotic resistance in diverse homologs of the dihydrofolate reductase protein family through broad mutational scanning.

ABSTRACT: Antimicrobial resistance studies often focus on individual protein variants, neglecting broader protein family dynamics. Dihydrofolate reductase (DHFR), a key antibiotic target, has been extensively studied using deep mutational scanning, yet resistance mechanisms across this diverse protein family remain poorly understood. Here, we developed a synthetic metagenomics approach using DropSynth, a scalable gene synthesis platform, to construct a phylogenetically diverse library of 1536 DHFR homologs. These sequences, primarily derived from host-associated metagenomes, represent 759 bacterial species, including many clinically relevant pathogens. A multiplexed in vivo assay tested their ability to restore metabolic function and confer trimethoprim resistance in an Escherichia coli ∆f

SUBMITTER: Romanowicz KJ 

PROVIDER: S-EPMC12346277 | biostudies-literature | 2025 Aug

REPOSITORIES: biostudies-literature