Conformational Rearrangements in the Sensory RcsF/OMP Complex Mediate Signal Transduction Across the Bacterial Cell Envelope

Overview

Journal PLoS Genet

Specialty Genetics

Date 2023 Jan 27

PMID 36706155

Authors

Sarah R Lach

Santosh Kumar

Seonghoon Kim

Wonpil Im

Anna Konovalova

Affiliations

Soon will be listed here.

Abstract

Timely detection and repair of envelope damage are paramount for bacterial survival. The Regulator of Capsule Synthesis (Rcs) stress response can transduce the stress signals across the multilayered gram-negative cell envelope to regulate gene expression in the cytoplasm. Previous studies defined the overall pathway, which begins with the sensory lipoprotein RcsF interacting with several outer membrane proteins (OMPs). RcsF can also interact with the periplasmic domain of the negative regulator IgaA, derepressing the downstream RcsCDB phosphorelay. However, how the RcsF/IgaA interaction is regulated at the molecular level to activate the signaling in response to stress remains poorly understood. In this study, we used a site-saturated mutant library of rcsF to carry out several independent genetic screens to interrogate the mechanism of signal transduction from RcsF to IgaA. We analyzed several distinct classes of rcsF signaling mutants, and determined the region of RcsF that is critically important for signal transduction. This region is bifunctional as it is important for RcsF interaction with both IgaA and OMPs. The mutant analysis provides strong evidence for conformational changes in the RcsF/OMP complex mediating signal transduction to IgaA, and the first direct evidence that OMPs play an important regulatory role in Rcs signaling.

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References

Umekawa M, Miyagawa H, Kondo D, Matsuoka S, Matsumoto K, Hara H . Importance of the proline-rich region for the regulatory function of RcsF, an outer membrane lipoprotein component of the Escherichia coli Rcs signal transduction system. Microbiology (Reading). 2013; 159(Pt 9):1818-1827. DOI: 10.1099/mic.0.069328-0. View

Rojas E, Billings G, Odermatt P, Auer G, Zhu L, Miguel A . The outer membrane is an essential load-bearing element in Gram-negative bacteria. Nature. 2018; 559(7715):617-621. PMC: 6089221. DOI: 10.1038/s41586-018-0344-3. View

Housden N, Wojdyla J, Korczynska J, Grishkovskaya I, Kirkpatrick N, Brzozowski A . Directed epitope delivery across the Escherichia coli outer membrane through the porin OmpF. Proc Natl Acad Sci U S A. 2010; 107(50):21412-7. PMC: 3003033. DOI: 10.1073/pnas.1010780107. View

Callewaert L, Vanoirbeek K, Lurquin I, Michiels C, Aertsen A . The Rcs two-component system regulates expression of lysozyme inhibitors and is induced by exposure to lysozyme. J Bacteriol. 2009; 191(6):1979-81. PMC: 2648352. DOI: 10.1128/JB.01549-08. View

Housden N, Hopper J, Lukoyanova N, Rodriguez-Larrea D, Wojdyla J, Klein A . Intrinsically disordered protein threads through the bacterial outer-membrane porin OmpF. Science. 2013; 340(6140):1570-4. PMC: 3856478. DOI: 10.1126/science.1237864. View

Wall E, Majdalani N, Gottesman S . IgaA negatively regulates the Rcs Phosphorelay via contact with the RcsD Phosphotransfer Protein. PLoS Genet. 2020; 16(7):e1008610. PMC: 7418988. DOI: 10.1371/journal.pgen.1008610. View

Meng J, Xu J, Huang C, Chen J . Rcs Phosphorelay Responses to Truncated Lipopolysaccharide-Induced Cell Envelope Stress in . Molecules. 2020; 25(23). PMC: 7730088. DOI: 10.3390/molecules25235718. View

Nikaido H . Molecular basis of bacterial outer membrane permeability revisited. Microbiol Mol Biol Rev. 2003; 67(4):593-656. PMC: 309051. DOI: 10.1128/MMBR.67.4.593-656.2003. View

Shiba Y, Miyagawa H, Nagahama H, Matsumoto K, Kondo D, Matsuoka S . Exploring the relationship between lipoprotein mislocalization and activation of the Rcs signal transduction system in Escherichia coli. Microbiology (Reading). 2012; 158(Pt 5):1238-1248. DOI: 10.1099/mic.0.056945-0. View

10.

Leverrier P, Declercq J, Denoncin K, Vertommen D, Hiniker A, Cho S . Crystal structure of the outer membrane protein RcsF, a new substrate for the periplasmic protein-disulfide isomerase DsbC. J Biol Chem. 2011; 286(19):16734-42. PMC: 3089515. DOI: 10.1074/jbc.M111.224865. View

11.

Tata M, Kumar S, Lach S, Saha S, Hart E, Konovalova A . High-throughput suppressor screen demonstrates that RcsF monitors outer membrane integrity and not Bam complex function. Proc Natl Acad Sci U S A. 2021; 118(32). PMC: 8364173. DOI: 10.1073/pnas.2100369118. View

12.

Asmar A, Ferreira J, Cohen E, Cho S, Beeby M, Hughes K . Communication across the bacterial cell envelope depends on the size of the periplasm. PLoS Biol. 2017; 15(12):e2004303. PMC: 5736177. DOI: 10.1371/journal.pbio.2004303. View

13.

Shiba Y, Yokoyama Y, Aono Y, Kiuchi T, Kusaka J, Matsumoto K . Activation of the Rcs signal transduction system is responsible for the thermosensitive growth defect of an Escherichia coli mutant lacking phosphatidylglycerol and cardiolipin. J Bacteriol. 2004; 186(19):6526-35. PMC: 516613. DOI: 10.1128/JB.186.19.6526-6535.2004. View

14.

Silhavy T, Kahne D, Walker S . The bacterial cell envelope. Cold Spring Harb Perspect Biol. 2010; 2(5):a000414. PMC: 2857177. DOI: 10.1101/cshperspect.a000414. View

15.

Tata M, Konovalova A . Improper Coordination of BamA and BamD Results in Bam Complex Jamming by a Lipoprotein Substrate. mBio. 2019; 10(3). PMC: 6529637. DOI: 10.1128/mBio.00660-19. View

16.

Mirdita M, Schutze K, Moriwaki Y, Heo L, Ovchinnikov S, Steinegger M . ColabFold: making protein folding accessible to all. Nat Methods. 2022; 19(6):679-682. PMC: 9184281. DOI: 10.1038/s41592-022-01488-1. View

17.

Benn G, Mikheyeva I, Inns P, Forster J, Ojkic N, Bortolini C . Phase separation in the outer membrane of . Proc Natl Acad Sci U S A. 2021; 118(44). PMC: 8612244. DOI: 10.1073/pnas.2112237118. View

18.

Dekoninck K, Letoquart J, Laguri C, Demange P, Bevernaegie R, Simorre J . Defining the function of OmpA in the Rcs stress response. Elife. 2020; 9. PMC: 7553776. DOI: 10.7554/eLife.60861. View

19.

Housden N, Loftus S, Moore G, James R, Kleanthous C . Cell entry mechanism of enzymatic bacterial colicins: porin recruitment and the thermodynamics of receptor binding. Proc Natl Acad Sci U S A. 2005; 102(39):13849-54. PMC: 1236540. DOI: 10.1073/pnas.0503567102. View

20.

Saha S, Lach S, Konovalova A . Homeostasis of the Gram-negative cell envelope. Curr Opin Microbiol. 2021; 61:99-106. PMC: 8577651. DOI: 10.1016/j.mib.2021.03.008. View