Abstract
Cell size, cell growth, and cell cycle events are necessarily intertwined to achieve robust bacterial replication. Yet, a comprehensive and integrated view of these fundamental processes is lacking. Here, we describe an image‐based quantitative screen of the single‐gene knockout collection of Escherichia coli and identify many new genes involved in cell morphogenesis, population growth, nucleoid (bulk chromosome) dynamics, and cell division. Functional analyses, together with high‐dimensional classification, unveil new associations of morphological and cell cycle phenotypes with specific functions and pathways. Additionally, correlation analysis across ~4,000 genetic perturbations shows that growth rate is surprisingly not predictive of cell size. Growth rate was also uncorrelated with the relative timings of nucleoid separation and cell constriction. Rather, our analysis identifies scaling relationships between cell size and nucleoid size and between nucleoid size and the relative timings of nucleoid separation and cell division. These connections suggest that the nucleoid links cell morphogenesis to the cell cycle.
Highlights
Cell size, cell growth, and cell cycle events are necessarily intertwined to achieve robust bacterial replication
We present a high-content, quantitative study that uses the Keio collection of Escherichia coli gene deletion strains (Baba et al, 2006) and combines microscopy with advanced statistical and image analysis procedures to examine the impact of each nonessential E. coli gene on cell morphology, cell size, growth, nucleoid dynamics, and cell constriction
The strains were grown in 96-well plates in M9 medium supplemented with 0.1% casamino acids and 0.2% glucose at 30°C
Summary
Cell growth, and cell cycle events are necessarily intertwined to achieve robust bacterial replication. We describe an image-based quantitative screen of the single-gene knockout collection of Escherichia coli and identify many new genes involved in cell morphogenesis, population growth, nucleoid (bulk chromosome) dynamics, and cell division. Correlation analysis across ~4,000 genetic perturbations shows that growth rate is surprisingly not predictive of cell size. Growth rate was uncorrelated with the relative timings of nucleoid separation and cell constriction. Our analysis identifies scaling relationships between cell size and nucleoid size and between nucleoid size and the relative timings of nucleoid separation and cell division. These connections suggest that the nucleoid links cell morphogenesis to the cell cycle
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