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Adaptation of the periplasm to maintain spatial constraints essential for cell envelope processes and cell viability

The cell envelope of Gram-negative bacteria consists of two membranes surrounding a periplasm and peptidoglycan layer. Molecular machines spanning the cell envelope depend on spatial constraints and load-bearing forces across the cell envelope and surface. The mechanisms dictating spatial constraint...

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Autores principales: Mandela, Eric, Stubenrauch, Christopher J, Ryoo, David, Hwang, Hyea, Cohen, Eli J, Torres, Von L, Deo, Pankaj, Webb, Chaille T, Huang, Cheng, Schittenhelm, Ralf B, Beeby, Morgan, Gumbart, JC, Lithgow, Trevor, Hay, Iain D
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8824477/
https://www.ncbi.nlm.nih.gov/pubmed/35084330
http://dx.doi.org/10.7554/eLife.73516
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author Mandela, Eric
Stubenrauch, Christopher J
Ryoo, David
Hwang, Hyea
Cohen, Eli J
Torres, Von L
Deo, Pankaj
Webb, Chaille T
Huang, Cheng
Schittenhelm, Ralf B
Beeby, Morgan
Gumbart, JC
Lithgow, Trevor
Hay, Iain D
author_facet Mandela, Eric
Stubenrauch, Christopher J
Ryoo, David
Hwang, Hyea
Cohen, Eli J
Torres, Von L
Deo, Pankaj
Webb, Chaille T
Huang, Cheng
Schittenhelm, Ralf B
Beeby, Morgan
Gumbart, JC
Lithgow, Trevor
Hay, Iain D
author_sort Mandela, Eric
collection PubMed
description The cell envelope of Gram-negative bacteria consists of two membranes surrounding a periplasm and peptidoglycan layer. Molecular machines spanning the cell envelope depend on spatial constraints and load-bearing forces across the cell envelope and surface. The mechanisms dictating spatial constraints across the cell envelope remain incompletely defined. In Escherichia coli, the coiled-coil lipoprotein Lpp contributes the only covalent linkage between the outer membrane and the underlying peptidoglycan layer. Using proteomics, molecular dynamics, and a synthetic lethal screen, we show that lengthening Lpp to the upper limit does not change the spatial constraint but is accommodated by other factors which thereby become essential for viability. Our findings demonstrate E. coli expressing elongated Lpp does not simply enlarge the periplasm in response, but the bacteria accommodate by a combination of tilting Lpp and reducing the amount of the covalent bridge. By genetic screening, we identified all of the genes in E. coli that become essential in order to enact this adaptation, and by quantitative proteomics discovered that very few proteins need to be up- or down-regulated in steady-state levels in order to accommodate the longer Lpp. We observed increased levels of factors determining cell stiffness, a decrease in membrane integrity, an increased membrane vesiculation and a dependance on otherwise non-essential tethers to maintain lipid transport and peptidoglycan biosynthesis. Further this has implications for understanding how spatial constraint across the envelope controls processes such as flagellum-driven motility, cellular signaling, and protein translocation
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spelling pubmed-88244772022-02-10 Adaptation of the periplasm to maintain spatial constraints essential for cell envelope processes and cell viability Mandela, Eric Stubenrauch, Christopher J Ryoo, David Hwang, Hyea Cohen, Eli J Torres, Von L Deo, Pankaj Webb, Chaille T Huang, Cheng Schittenhelm, Ralf B Beeby, Morgan Gumbart, JC Lithgow, Trevor Hay, Iain D eLife Cell Biology The cell envelope of Gram-negative bacteria consists of two membranes surrounding a periplasm and peptidoglycan layer. Molecular machines spanning the cell envelope depend on spatial constraints and load-bearing forces across the cell envelope and surface. The mechanisms dictating spatial constraints across the cell envelope remain incompletely defined. In Escherichia coli, the coiled-coil lipoprotein Lpp contributes the only covalent linkage between the outer membrane and the underlying peptidoglycan layer. Using proteomics, molecular dynamics, and a synthetic lethal screen, we show that lengthening Lpp to the upper limit does not change the spatial constraint but is accommodated by other factors which thereby become essential for viability. Our findings demonstrate E. coli expressing elongated Lpp does not simply enlarge the periplasm in response, but the bacteria accommodate by a combination of tilting Lpp and reducing the amount of the covalent bridge. By genetic screening, we identified all of the genes in E. coli that become essential in order to enact this adaptation, and by quantitative proteomics discovered that very few proteins need to be up- or down-regulated in steady-state levels in order to accommodate the longer Lpp. We observed increased levels of factors determining cell stiffness, a decrease in membrane integrity, an increased membrane vesiculation and a dependance on otherwise non-essential tethers to maintain lipid transport and peptidoglycan biosynthesis. Further this has implications for understanding how spatial constraint across the envelope controls processes such as flagellum-driven motility, cellular signaling, and protein translocation eLife Sciences Publications, Ltd 2022-01-27 /pmc/articles/PMC8824477/ /pubmed/35084330 http://dx.doi.org/10.7554/eLife.73516 Text en © 2022, Mandela et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Cell Biology
Mandela, Eric
Stubenrauch, Christopher J
Ryoo, David
Hwang, Hyea
Cohen, Eli J
Torres, Von L
Deo, Pankaj
Webb, Chaille T
Huang, Cheng
Schittenhelm, Ralf B
Beeby, Morgan
Gumbart, JC
Lithgow, Trevor
Hay, Iain D
Adaptation of the periplasm to maintain spatial constraints essential for cell envelope processes and cell viability
title Adaptation of the periplasm to maintain spatial constraints essential for cell envelope processes and cell viability
title_full Adaptation of the periplasm to maintain spatial constraints essential for cell envelope processes and cell viability
title_fullStr Adaptation of the periplasm to maintain spatial constraints essential for cell envelope processes and cell viability
title_full_unstemmed Adaptation of the periplasm to maintain spatial constraints essential for cell envelope processes and cell viability
title_short Adaptation of the periplasm to maintain spatial constraints essential for cell envelope processes and cell viability
title_sort adaptation of the periplasm to maintain spatial constraints essential for cell envelope processes and cell viability
topic Cell Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8824477/
https://www.ncbi.nlm.nih.gov/pubmed/35084330
http://dx.doi.org/10.7554/eLife.73516
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