Bacteria Getting into Shape: Genetic Determinants of E. coli Morphology
| Autor: | Shawn French, Jean-Philippe Côtôé, Jonathan M. Stokes, Ray Truant, Eric D. Brown, Paul Dunman |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
030106 microbiology Mutant Observation Computational biology Biology medicine.disease_cause Cell morphology Microbiology 03 medical and health sciences Interaction network Virology medicine Genomic library Gene Regulatory Networks Genetic Testing Escherichia coli Gene Gene knockout Microscopy Escherichia coli K12 Phenotype QR1-502 030104 developmental biology Genes Bacterial Gene Deletion |
| Zdroj: | mBio, Vol 8, Iss 2 (2017) mBio mBio, Vol 8, Iss 2, p e01977-16 (2017) |
| ISSN: | 2150-7511 |
| DOI: | 10.1128/mBio.01977-16 |
| Popis: | Perturbation of cellular processes is a prevailing approach to understanding biology. To better understand the complicated biology that defines bacterial shape, a sensitive, high-content platform was developed to detect multiple morphological defect phenotypes using microscopy. We examined morphological phenotypes across the Escherichia coli K-12 deletion (Keio) collection at the mid-exponential growth phase, revealing 111 deletions perturbing shape. Interestingly, 64% of these were uncharacterized mutants, illustrating the complex nature of shape maintenance and regulation in bacteria. To understand the roles these genes play in defining morphology, 53 mutants with knockouts resulting in abnormal cell shape were crossed with the Keio collection in high throughput, generating 1,373 synthetic lethal interactions across 1.7 million double deletion mutants. This analysis yielded a highly populated interaction network spanning and linking multiple phenotypes, with a preponderance of interactions involved in transport, oxidation-reduction, and metabolic processes. IMPORTANCE Genetic perturbations of cellular functions are a prevailing approach to understanding cell systems, which are increasingly being practiced in very high throughput. Here, we report a high-content microscopy platform tailored to bacteria, which probes the impact of genetic mutation on cell morphology. This has particular utility in revealing elusive and subtle morphological phenotypes associated with blocks in nonessential cellular functions. We report 111 nonessential mutations impacting E. coli morphology, with nearly half of those genes being poorly annotated or uncharacterized. Further, these genes appear to be tightly linked to transport or redox processes within the cell. The screening platform is simple and low cost and is broadly applicable to any bacterial genomic library or chemical collection. Indeed, this is a powerful tool in understanding the biology behind bacterial shape. |
| Databáze: | OpenAIRE |
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