Structural mechanism of a DNA polymerase critical for developing antibiotic-resistance

Structural mechanism of a DNA polymerase critical for developing antibiotic-resistanceAbstractAntibiotic resistance is a dire and growing threat to human health, with over 20000 deaths per year occurring inthe US alone due to drug-resistant pathogenic bacteria. Unfortunately, current antibiotics only target a fewcellular pathways, leading to widespread emergence of multi-drug resistant strains. Moreover, development ofnew antibiotics has largely stalled. Here we propose investigation of DnaE2 as an exciting target fordevelopment of novel antibiotics that minimize evolution of drug resistance. DnaE2?s error-prone DNApolymerase activity is important for evolving drug resistance across a wide spectrum of bacteria. Therefore, aDnaE2 inhibitor could be used in combination therapy to prevent emergence of drug resistance. Furthermore,DnaE2 is critical for pathogenesis in Mycobacterium tuberculosis, but is not necessary for bacterial growth.Thus, DnaE2 inhibitors would prevent disease without selecting for drug-resistant bacterial strains. Finally,DnaE2 is unrelated to eukaryotic DNA polymerases, minimizing the chance of cross-reaction of DnaE2inhibitors with human enzymes. These advantages make DnaE2 a particularly exciting new antibiotic target.Our expertise in structural, biophysical, and biochemical studies of DNA polymerases and other replicationproteins uniquely positions the Kelch lab to investigate DnaE2?s potential as a drug target. We will determinethe first atomic structure of DnaE2, which will reveal the molecular basis for DnaE2?s mutagenic activity as wellas provide a blueprint for developing potent small molecule inhibitors. We will also measure the mutagenicpreference for DnaE2, which will reveal how the enzyme drives the evolution of antibiotic resistance. Finally,we will screen for small molecule inhibitors of DnaE2, which will provide the foundation for developing novel?anti-evolution? antibiotics.