Appropriate Regulation of the σ^E-Dependent Envelope Stress Response Is Necessary To Maintain Cell Envelope Integrity and Stationary-Phase Survival in Escherichia coli.

The alternative sigma factor σ^E is a key component of the Escherichia coli response to cell envelope stress and is required for viability even in the absence of stress. The activity of σ^E increases during entry into stationary phase, suggesting an important role for σ^E when nutrients are limiting. Elevated σ^E activity has been proposed to activate a pathway leading to the lysis of nonculturable cells that accumulate during early stationary phase. To better understand σ^E-directed cell lysis and the role of σ^E in stationary phase, we investigated the effects of elevated σ^E activity in cultures grown for 10 days. We demonstrate that high σ^E activity is lethal for all cells in stationary phase, not only those that are nonculturable. Spontaneous mutants with reduced σ^E activity, due primarily to point mutations in the region of σ^E that binds the -35 promoter motif, arise and take over cultures within 5 to 6 days after entry into stationary phase. High σ^E activity leads to large reductions in the levels of outer membrane porins and increased membrane permeability, indicating membrane defects. These defects can be counteracted and stationary-phase lethality delayed significantly by stabilizing membranes with Mg²⁺ and buffering the growth medium or by deleting the σ^E-dependent small RNAs (sRNAs) MicA, RybB, and MicL, which inhibit the expression of porins and Lpp. Expression of these sRNAs also reverses the loss of viability following depletion of σ^E activity. Our results demonstrate that appropriate regulation of σ^E activity, ensuring that it is neither too high nor too low, is critical for envelope integrity and cell viability.IMPORTANCE The Gram-negative cell envelope and cytoplasm differ significantly, and separate responses have evolved to combat stress in each compartment. An array of cell envelope stress responses exist, each of which is focused on different parts of the envelope. The σ^E response is conserved in many enterobacteria and is tuned to monitor pathways for the maturation and delivery of outer membrane porins, lipoproteins, and lipopolysaccharide to the outer membrane. The activity of σ^E is tightly regulated to match the production of σ^E regulon members to the needs of the cell. In E. coli, loss of σ^E results in lethality. Here we demonstrate that excessive σ^E activity is also lethal and results in decreased membrane integrity, the very phenotype the system is designed to prevent.

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