Exploring Antibiotic Resistance in Diverse Homologs of the Dihydrofolate Reductase Protein Family through Broad Mutational Scanning.

ABSTRACT: Current antibiotic 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. Using DropSynth, a scalable gene synthesis platform, we designed a library of 1,536 synthetic DHFR homologs representing 778 species of bacteria, archaea, and viruses, including clinically relevant pathogens. A multiplexed in vivo assay tested their ability to restore metabolic function and confer trimethoprim resistance in an E. coli ∆folA strain. Over half of the synthetic homologs rescued the phenotype without supplementation, and mutants with up t

SUBMITTER: Romanowicz KJ 

PROVIDER: S-EPMC11785229 | biostudies-literature | 2025 Jan

REPOSITORIES: biostudies-literature