Abstract
Bacteria survive in the environment with three steps: a sensing environmental conditions, a responding to sensed signals, and an adaptation for proper survival in the environment. An adapting bacterial cell occurs cell division to increase the number of sister cells, termed adaptive growth. Two-component systems (TCSs), representing the main bacterial signal transduction systems, consist of a pair of one sensor kinase (SK) and one response regulator (RR), and RR genes are abundant in most bacterial genomes as part of the core genome. The OmpR gene family, a group of RR genes, is conserved in 95% of known bacterial genomes. The Escherichia coli genome has an estimated 34 RR genes in total, including 14 genes of OmpR family genes. To reveal the contribution of TCSs for fast growth as an adaptive growth strategy of E. coli, we isolated a set of gene knockout strains by using newly developed genome editing technology, the HoSeI (Homologous Sequence Integration) method, based on CRISPR-Cas9. The statistics of single cell observation show a knockout of an arbitrary pair of phoP, phoB, and ompR genes, stably expressed by positive feedback regulation, dramatically inhibit the optimum adaptive growth of E. coli. These insights suggest that the adaptive growth of bacteria is fulfilled by the optimum high intracellular level of regulators acquired during growth under environmental conditions.
Highlights
Bacteria survive and increase their population by binary cell division in a timely manner
To gain insight into the effect of RR genes on growth, we isolated the set of all combinations of single, double, and triple-gene knockout strains by using the newly developed Homologous sequence integration (HoSeI) method for the E. coli genome based on CRISPR-Cas
Bacterial genomes less than 5 Mbp in size contained the relative RR Clusters of Orthologous Groups of proteins (COGs) number of 2 RR COGs per 1 Mbp of genome, while bacterial genomes more than 5 Mbp in size contained 8 RR COGs (Fig. 1A)
Summary
Bacteria survive and increase their population by binary cell division in a timely manner. We knocked out a pair of similar genes and isolated four double RR knockout strains of Δ phoP Δ kdpE, Δ phoB Δ creB, Δ ompR Δ cpxR, and Δ ompR Δ rstA (see above) (Fig. S4).
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