Supramolecular assemblies underpin turnover of outer membrane proteins in bacteria

DC FieldValueLanguage
dc.contributor.authorRassam, Patrice
dc.contributor.authorCopeland, Nikki A.
dc.contributor.authorBirkholz, Oliver
dc.contributor.authorToth, Csaba
dc.contributor.authorChavent, Matthieu
dc.contributor.authorDuncan, Anna L.
dc.contributor.authorCross, Stephen J.
dc.contributor.authorHousden, Nicholas G.
dc.contributor.authorKaminska, Renata
dc.contributor.authorSeger, Urban
dc.contributor.authorQuinn, Diana M.
dc.contributor.authorGarrod, Tamsin J.
dc.contributor.authorSansom, Mark S. P.
dc.contributor.authorPiehler, Jacob
dc.contributor.authorBaumann, Christoph G.
dc.contributor.authorKleanthous, Colin
dc.date.accessioned2021-12-23T16:16:18Z-
dc.date.available2021-12-23T16:16:18Z-
dc.date.issued2015
dc.identifier.issn00280836
dc.identifier.urihttps://osnascholar.ub.uni-osnabrueck.de/handle/unios/11808-
dc.description.abstractGram-negative bacteria inhabit a broad range of ecological niches. For Escherichia coli, this includes river water as well as humans and animals, where it can be both a commensal and a pathogen(1-3). Intricate regulatory mechanisms ensure that bacteria have the right complement of beta-barrel outer membrane proteins (OMPs) to enable adaptation to a particular habitat(4,5). Yet no mechanism is known for replacing OMPs in the outer membrane, an issue that is further confounded by the lack of an energy source and the high stability(6) and abundance of OMPs(5). Here we uncover the process underpinning OMP turnover in Escherichia coli and show it to be passive and binary in nature, in which old OMPs are displaced to the poles of growing cells as new OMPs take their place. Using fluorescent colicins as OMP-specific probes, in combination with ensemble and single-molecule fluorescence microscopy in vivo and in vitro, as well as molecular dynamics simulations, we established the mechanism for binary OMP partitioning. OMPs clustered to form 0.5-mu m diameter islands, where their diffusion is restricted by promiscuous interactions with other OMPs. OMP islands were distributed throughout the cell and contained the Bam complex, which catalyses the insertion of OMPs in the outer membrane(7,8). However, OMP biogenesis occurred as a gradient that was highest at mid-cell but largely absent at cell poles. The cumulative effect is to push old OMP islands towards the poles of growing cells, leading to a binary distribution when cells divide. Hence, the outer membrane of a Gram-negative bacterium is a spatially and temporally organized structure, and this organization lies at the heart of how OMPs are turned over in the membrane.
dc.description.sponsorshipRoyal SocietyRoyal Society of LondonEuropean Commission [2004/R1]; University of York (Research Priming Fund); BBSRC LoLa grantUK Research & Innovation (UKRI)Biotechnology and Biological Sciences Research Council (BBSRC) [BB/G020671/1]; Deutsche ForschungsgemeinschaftGerman Research Foundation (DFG) [SFB 944]; BBSRCUK Research & Innovation (UKRI)Biotechnology and Biological Sciences Research Council (BBSRC) [BB/L002558/1]; Wellcome TrustWellcome TrustEuropean Commission [WT092970MA]; Biotechnology and Biological Sciences Research CouncilUK Research & Innovation (UKRI)Biotechnology and Biological Sciences Research Council (BBSRC) [BB/L002558/1, BB/H000267/1, BB/G020671/2, BB/I019855/1, BEP17032, B19456, BBS/B/16011] Funding Source: researchfish; Engineering and Physical Sciences Research CouncilUK Research & Innovation (UKRI)Engineering & Physical Sciences Research Council (EPSRC) [EP/J010421/1, EP/L000253/1] Funding Source: researchfish; BBSRCUK Research & Innovation (UKRI)Biotechnology and Biological Sciences Research Council (BBSRC) [BB/H000267/1, BB/G020671/2, BB/I019855/1] Funding Source: UKRI; EPSRCUK Research & Innovation (UKRI)Engineering & Physical Sciences Research Council (EPSRC) [EP/L000253/1, EP/J010421/1] Funding Source: UKRI; We thank the University of Oxford Micron facility and the University of York Biology Technology Facility for access to microscopy facilities. The authors also thank S. Buchanan and K. Jakes for anti-BamC antibody and Colla plasmids, I. Henderson for HA-tagged BamA constructs, C. Robinson for TatA-GFP construct, K. Heurlier and J. Pullen for PCR screening of deletion strains, A. Sharma for the design of the inactivating disulfide bond in ColIa, and P. E. Milhier and his group for providing optimised PATRACK single-molecule tracking software. A.L.D. and M.C. thank T. Reddy for discussions and assistance with diffusion calculations. U.S. and C.G.B. thank the University of York Biology Electronic and Mechanical Workshops for custom fabrications, and G. Mashanov for custom image acquisition software. We also thank B. Berks for comments on the manuscript. C.G.B. acknowledges the Royal Society (2004/R1) and the University of York (Research Priming Fund) for financial support used to develop the TIRFM. P.R. acknowledges the late R. Saadia for his unwavering support. M.S.P.S. acknowledges access to the UK supercomputer ARCHER for molecular dynamics simulations. This work was supported by grants to C.K. and C.G.B. (BBSRC LoLa grant BB/G020671/1), J.P. (Deutsche Forschungsgemeinschaft SFB 944) and M.S.P.S. (BBSRC BB/L002558/1, Wellcome Trust WT092970MA).
dc.language.isoen
dc.publisherNATURE PUBLISHING GROUP
dc.relation.ispartofNATURE
dc.subjectCELLS
dc.subjectDIFFUSION
dc.subjectDYNAMICS
dc.subjectESCHERICHIA-COLI
dc.subjectINSERTION
dc.subjectLOCALIZE PROTEINS
dc.subjectMultidisciplinary Sciences
dc.subjectPOLYMER-SUPPORTED MEMBRANES
dc.subjectR-DOMAIN
dc.subjectRECEPTOR-BINDING
dc.subjectScience & Technology - Other Topics
dc.subjectSINGLE-MOLECULE
dc.titleSupramolecular assemblies underpin turnover of outer membrane proteins in bacteria
dc.typejournal article
dc.identifier.doi10.1038/nature14461
dc.identifier.isiISI:000357950900039
dc.description.volume523
dc.description.issue7560
dc.description.startpage333+
dc.contributor.orcid0000-0001-6360-7959
dc.contributor.orcid0000-0003-4524-4773
dc.contributor.orcid0000-0001-9873-4552
dc.contributor.orcid0000-0003-3565-0479
dc.contributor.orcid0000-0002-8630-0957
dc.contributor.orcid0000-0001-6551-3219
dc.contributor.orcid0000-0002-6204-4717
dc.contributor.researcheridAAM-9860-2020
dc.contributor.researcheridC-5157-2009
dc.contributor.researcheridP-6564-2014
dc.identifier.eissn14764687
dc.publisher.placeMACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND
dcterms.isPartOf.abbreviationNature
dcterms.oaStatusGreen Accepted
crisitem.author.deptFB 05 - Biologie/Chemie-
crisitem.author.deptidfb05-
crisitem.author.orcid0000-0002-2143-2270-
crisitem.author.parentorgUniversität Osnabrück-
crisitem.author.netidPiJa938-
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