Bacteria Getting into Shape: Genetic Determinants of E. coli Morphology
Bacteria Getting into Shape: Genetic Determinants of E. coli Morphology
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DOI：
10.1128/mbio.01977-16
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发表时间：
2017-03-01
期刊：
影响因子：
6.4
通讯作者：
Brown, Eric D.
中科院分区：
文献类型：
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作者：
French, Shawn;Cote, Jean-Philippe;Brown, Eric D.
Perturbation of cellular processes is a prevailing approach to understanding biology.为了更好地了解定义细菌形状的复杂生物学，开发了一个灵敏的高内涵平台，用于使用显微镜检测多种形态缺陷表型。我们检查了指数生长期中大肠杆菌 K-12 缺失 (Keio) 集合的形态表型，揭示了 111 个缺失扰乱形状。有趣的是，其中 64% 是未表征的突变体，这说明了细菌形状维持和调节的复杂性。为了了解这些基因在定义形态学中所起的作用，将 53 个敲除导致细胞形状异常的突变体与 Keio 系列进行高通量杂交，在 170 万个双缺失突变体中产生了 1,373 个合成致死相互作用。该分析产生了一个跨越和连接多种表型的高度密集的相互作用网络，其中大部分相互作用涉及运输、氧化还原和代谢过程。 重要性 细胞功能的遗传扰动是理解细胞系统的流行方法，这种方法越来越多地以非常高的通量进行实践。 Here, we report a high-content microscopy platform tailored to bacteria, which probes the impact of genetic mutation on cell morphology.这对于揭示与非必需细胞功能障碍相关的难以捉摸和微妙的形态表型特别有用。 We report 111 nonessential mutations impacting E. coli morphology, with nearly half of those genes being poorly annotated or uncharacterized.此外，这些基因似乎与细胞内的运输或氧化还原过程紧密相关。该筛选平台简单、成本低，广泛适用于任何细菌基因组文库或化学保藏。 Indeed, this is a powerful tool in understanding the biology behind bacterial shape.
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.