Revealing bacterial targets of growth inhibitors encoded by bacteriophage T7
| Autor: | Michael Gershovits, Ruth Kiro, Miriam Manor, Shahar Molshanski-Mor, Rotem Edgar, Udi Qimron, Tal Pupko, Ido Yosef, Mia Laserson |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Mutant
Genome Viral MreB Microbiology Gene product Bacteriophage 03 medical and health sciences chemistry.chemical_compound Viral Proteins Bacteriophage T7 Escherichia coli Cytoskeleton Gene 030304 developmental biology 0303 health sciences Multidisciplinary biology 030306 microbiology Escherichia coli Proteins Polynucleotide Adenylyltransferase Biological Sciences biology.organism_classification chemistry DNA Viral Growth inhibition Bacteria |
| Popis: | Today's arsenal of antibiotics is ineffective against some emerging strains of antibiotic-resistant pathogens. Novel inhibitors of bacterial growth therefore need to be found. The target of such bacterial-growth inhibitors must be identified, and one way to achieve this is by locating mutations that suppress their inhibitory effect. Here, we identified five growth inhibitors encoded by T7 bacteriophage. High-throughput sequencing of genomic DNA of resistant bacterial mutants evolving against three of these inhibitors revealed unique mutations in three specific genes. We found that a nonessential host gene, ppiB, is required for growth inhibition by one bacteriophage inhibitor and another nonessential gene, pcnB, is required for growth inhibition by a different inhibitor. Notably, we found a previously unidentified growth inhibitor, gene product (Gp) 0.6, that interacts with the essential cytoskeleton protein MreB and inhibits its function. We further identified mutations in two distinct regions in the mreB gene that overcome this inhibition. Bacterial two-hybrid assay and accumulation of Gp0.6 only in MreB-expressing bacteria confirmed interaction of MreB and Gp0.6. Expression of Gp0.6 resulted in lemon-shaped bacteria followed by cell lysis, as previously reported for MreB inhibitors. The described approach may be extended for the identification of new growth inhibitors and their targets across bacterial species and in higher organisms. |
| Databáze: | OpenAIRE |
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