Competitive Fitness of Essential Gene Knockdowns Reveals a Broad-Spectrum Antibacterial Inhibitor of the Cell Division Protein FtsZ
Autor: | Xuan Li, Olga Riabova, Michael Domaratzki, Laurent R. Chiarelli, Silvia Buroni, Silvia T. Cardona, Marco Biggiogera, Giovanna Riccardi, Natalia Monakhova, Alberto Azzalin, April S. Gislason, Maria S. Stietz, Andrew M. Hogan, Viola Camilla Scoffone, Ann Karen C. Brassinga, Haben H Tesfu, Vadim Makarov |
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Rok vydání: | 2018 |
Předmět: |
0301 basic medicine
Cystic Fibrosis Burkholderia cenocepacia Cell division Burkholderia Green Fluorescent Proteins 030106 microbiology Mutant Drug Evaluation Preclinical Microbial Sensitivity Tests FtsZ fluorescent image analysis Green fluorescent protein 03 medical and health sciences Bacterial Proteins drug targets Animals Humans essential genes Pharmacology (medical) Caenorhabditis elegans Mechanisms of Action: Physiological Effects Illumina dye sequencing Pharmacology Genes Essential biology Tn-seq High-Throughput Nucleotide Sequencing Burkholderia Infections biology.organism_classification Anti-Bacterial Agents Cell biology mechanisms of action Cytoskeletal Proteins Infectious Diseases Essential gene Gene Knockdown Techniques Mutation biology.protein |
Zdroj: | Antimicrobial Agents and Chemotherapy |
ISSN: | 1098-6596 0066-4804 |
Popis: | To streamline the elucidation of antibacterial compounds’ mechanism of action, comprehensive high-throughput assays interrogating multiple putative targets are necessary. However, current chemogenomic approaches for antibiotic target identification have not fully utilized the multiplexing potential of next-generation sequencing. To streamline the elucidation of antibacterial compounds’ mechanism of action, comprehensive high-throughput assays interrogating multiple putative targets are necessary. However, current chemogenomic approaches for antibiotic target identification have not fully utilized the multiplexing potential of next-generation sequencing. Here, we used Illumina sequencing of transposon insertions to track the competitive fitness of a Burkholderia cenocepacia library containing essential gene knockdowns. Using this method, we characterized a novel benzothiadiazole derivative, 10126109 (C109), with antibacterial activity against B. cenocepacia, for which whole-genome sequencing of low-frequency spontaneous drug-resistant mutants had failed to identify the drug target. By combining the identification of hypersusceptible mutants and morphology screening, we show that C109 targets cell division. Furthermore, fluorescence microscopy of bacteria harboring green fluorescent protein (GFP) cell division protein fusions revealed that C109 prevents divisome formation by altering the localization of the essential cell division protein FtsZ. In agreement with this, C109 inhibited both the GTPase and polymerization activities of purified B. cenocepacia FtsZ. C109 displayed antibacterial activity against Gram-positive and Gram-negative cystic fibrosis pathogens, including Mycobacterium abscessus. C109 effectively cleared B. cenocepacia infection in the Caenorhabditis elegans model and exhibited additive interactions with clinically relevant antibiotics. Hence, C109 is an enticing candidate for further drug development. |
Databáze: | OpenAIRE |
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