Bacterial cell widening alters periplasmic size and activates envelope stress responses

Zietek, Matylda and Miguel, Amanda and Shi, Handuo and Khusainov, Iskander and Asmar, Abir T and Ram, Sri and Wartel, Morgane and Sueki, Anna and Schorb, Martin and Goulian, Mark and Collet, Jean-François and Beck, Martin and Huang, Kerwyn Casey and Typas, Athanasios (2025) Bacterial cell widening alters periplasmic size and activates envelope stress responses. The EMBO Journal, 44 (20). pp. 5816-5833. ISSN 1460-2075

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Abstract

The Rcs signal transduction system is a phosphorelay responsible for sensing enterobacterial cell envelope stresses. In Escherichia coli , the Rcs system is required to survive treatment with A22 and mecillinam, antibiotics that perturb cell size. To test whether size changes are correlated with envelope damage and thereby sensed by the Rcs system, we tuned E. coli cell size via A22 treatment, mutations in the cell-shape determinant MreB, and mechanically confined growth. In all conditions, cell width was strongly correlated with Rcs activation, and RcsF, the outer-membrane-localized upstream component, was essential for responding to cell width changes. Several gene deletions that induce Rcs resulted in cells that were wider than wild-type. Cryo-electron microscopy revealed that the periplasm of a wide MreB mutant is ~3 nm thinner than in wild-type cells, bringing RcsF closer to the downstream, inner-membrane-localized components of the signaling cascade. Conversely, extending the RcsF linker region in wild-type cells by ~3 nm increased Rcs activity. Thus, we propose that the Rcs system responds to changes in cell width due to altered periplasmic thickness.

Item Type:	Article
Authors:	Zietek, Matylda and Miguel, Amanda and Shi, Handuo and Khusainov, Iskander and Asmar, Abir T and Ram, Sri and Wartel, Morgane and Sueki, Anna and Schorb, Martin and Goulian, Mark and Collet, Jean-François and Beck, Martin and Huang, Kerwyn Casey and Typas, Athanasios
Document Language:	Language English
Subjects:	Natural Sciences > Life sciences; biology > Physiology & related subjects Natural Sciences > Life sciences; biology > Biochemistry Natural Sciences > Life sciences; biology > Microorganisms, fungi & algae
Divisions:	Azim Premji University - Bengaluru > School of Arts and Sciences
Full Text Status:	Public
URI:	http://publications.azimpremjiuniversity.edu.in/id/eprint/6935
Publisher URL:	https://doi.org/10.1038/s44318-025-00534-w

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