1. Elucidation of DnaE as the Antibacterial Target of the Natural Product, Nargenicin
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M A Powles, David B. Olsen, Katherine Young, Li Xiao, Nicholas Murgolo, Karen Dorso, Edward DiNunzio, Fred Racine, Gregory C. Adam, Douglas Wisniewski, Michael A. Goetz, Nichelle Hairston, Marta Arocho, Jing Su, Olga Genilloud, Ronald E. Painter, Robert G. K. Donald, Charles Gill, Bakela Nare, Francisca Vicente, and Milton L. Hammond more...
- Subjects
DNA Replication ,Staphylococcus aureus ,dnaE ,Topoisomerase IV ,DNA polymerase ,Mutant ,Clinical Biochemistry ,DNA-Directed DNA Polymerase ,medicine.disease_cause ,DNA gyrase ,Biochemistry ,chemistry.chemical_compound ,Inhibitory Concentration 50 ,Lactones ,Drug Discovery ,Drug Resistance, Bacterial ,medicine ,Escherichia coli ,Molecular Biology ,DNA Polymerase III ,Nucleic Acid Synthesis Inhibitors ,Pharmacology ,Nargenicin ,biology ,General Medicine ,Molecular biology ,Anti-Bacterial Agents ,chemistry ,Mutation ,biology.protein ,Molecular Medicine ,Efflux - Abstract
SummaryResistance to existing classes of antibiotics drives the need for discovery of novel compounds with unique mechanisms of action. Nargenicin A1, a natural product with limited antibacterial spectrum, was rediscovered in a whole-cell antisense assay. Macromolecular labeling in both Staphylococcus aureus and an Escherichia coli tolC efflux mutant revealed selective inhibition of DNA replication not due to gyrase or topoisomerase IV inhibition. S. aureus nargenicin-resistant mutants were selected at a frequency of ∼1 × 10−9, and whole-genome resequencing found a single base-pair change in the dnaE gene, a homolog of the E. coli holoenzyme α subunit. A DnaE single-enzyme assay was exquisitely sensitive to inhibition by nargenicin, and other in vitro characterization studies corroborated DnaE as the target. Medicinal chemistry efforts may expand the spectrum of this novel mechanism antibiotic. more...
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