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2021-12-27Zeitschriftenartikel DOI: 10.18452/26144
BldD-based bimolecular fluorescence complementation for in vivo detection of the second messenger cyclic di-GMP
dc.contributor.authorHalte, Manuel
dc.contributor.authorWörmann, Mirka E.
dc.contributor.authorBogisch, Maxim
dc.contributor.authorErhardt, Marc
dc.contributor.authorTschowri, Natalia
dc.date.accessioned2023-02-28T11:38:20Z
dc.date.available2023-02-28T11:38:20Z
dc.date.issued2021-12-27none
dc.identifier.urihttp://edoc.hu-berlin.de/18452/26806
dc.description.abstractThe widespread bacterial second messenger bis-(3′-5′)-cyclic diguanosine monophosphate (c-di-GMP) is an important regulator of biofilm formation, virulence and cell differentiation. C-di-GMP-specific biosensors that allow detection and visualization of c-di-GMP levels in living cells are key to our understanding of how c-di-GMP fluctuations drive cellular responses. Here, we describe a novel c-di-GMP biosensor, CensYBL, that is based on c-di-GMP-induced dimerization of the effector protein BldD from Streptomyces resulting in bimolecular fluorescence complementation of split-YPet fusion proteins. As a proof-of-principle, we demonstrate that CensYBL is functional in detecting fluctuations in intracellular c-di-GMP levels in the Gram-negative model bacteria Escherichia coli and Salmonella enterica serovar Typhimurium. Using deletion mutants of c-di-GMP diguanylate cyclases and phosphodiesterases, we show that c-di-GMP dependent dimerization of CBldD-YPet results in fluorescence complementation reflecting intracellular c-di-GMP levels. Overall, we demonstrate that the CensYBL biosensor is a user-friendly and versatile tool that allows to investigate c-di-GMP variations using single-cell and population-wide experimental set-ups.eng
dc.language.isoengnone
dc.publisherHumboldt-Universität zu Berlin
dc.rights(CC BY-NC-ND 4.0) Attribution-NonCommercial-NoDerivatives 4.0 Internationalger
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subjectbimolecular fluorescence complementationeng
dc.subjectbiosensoreng
dc.subjectBldDeng
dc.subjectc-di-GMPeng
dc.subjectdiguanylate cyclaseeng
dc.subjectEALeng
dc.subjectGGDEFeng
dc.subjectphosphodiesteraseeng
dc.subject.ddc570 Biologienone
dc.titleBldD-based bimolecular fluorescence complementation for in vivo detection of the second messenger cyclic di-GMPnone
dc.typearticle
dc.identifier.urnurn:nbn:de:kobv:11-110-18452/26806-3
dc.identifier.doihttp://dx.doi.org/10.18452/26144
dc.type.versionpublishedVersionnone
local.edoc.pages9none
local.edoc.type-nameZeitschriftenartikel
local.edoc.container-typeperiodical
local.edoc.container-type-nameZeitschrift
dc.description.versionPeer Reviewednone
dc.identifier.eissn1365-2958
dcterms.bibliographicCitation.doi10.1111/mmi.14876
dcterms.bibliographicCitation.journaltitleMolecular microbiologynone
dcterms.bibliographicCitation.volume117none
dcterms.bibliographicCitation.issue3none
dcterms.bibliographicCitation.originalpublishernameWiley-Blackwellnone
dcterms.bibliographicCitation.originalpublisherplaceOxford [u.a.]none
dcterms.bibliographicCitation.pagestart705none
dcterms.bibliographicCitation.pageend713none
bua.departmentLebenswissenschaftliche Fakultätnone

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