Abstract
ABSTRACTPerturbation 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.
Highlights
Perturbation of cellular processes is a prevailing approach to understanding biology
We report a high-content microscopy platform tailored to bacteria, which probes the impact of genetic mutation on cell morphology
The screening platform is simple and low cost and is broadly applicable to any bacterial genomic library or chemical collection. This is a powerful tool in understanding the biology behind bacterial shape
Summary
Perturbation of cellular processes is a prevailing approach to understanding biology. We report a high-content microscopy platform tailored to bacteria, which probes the impact of genetic mutation on cell morphology. These genes appear to be tightly linked to transport or redox processes within the cell.
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