Visualizing the dynamics of exported bacterial proteins with the chemogenetic fluorescent reporter FAST

Abstract Bacterial proteins exported to the cell surface play key cellular functions. However, despite the interest to study the localisation of surface proteins such as adhesins, transporters or hydrolases, monitoring their dynamics in live imaging remains challenging, due to the limited availabili...

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Autores principales:	Yankel Chekli, Caroline Peron-Cane, Dario Dell’Arciprete, Jean-François Allemand, Chenge Li, Jean-Marc Ghigo, Arnaud Gautier, Alice Lebreton, Nicolas Desprat, Christophe Beloin
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/74bfb02b768543aba2336c13542387e2
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spelling	oai:doaj.org-article:74bfb02b768543aba2336c13542387e22021-12-02T18:48:08ZVisualizing the dynamics of exported bacterial proteins with the chemogenetic fluorescent reporter FAST10.1038/s41598-020-72498-22045-2322https://doaj.org/article/74bfb02b768543aba2336c13542387e22020-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-72498-2https://doaj.org/toc/2045-2322Abstract Bacterial proteins exported to the cell surface play key cellular functions. However, despite the interest to study the localisation of surface proteins such as adhesins, transporters or hydrolases, monitoring their dynamics in live imaging remains challenging, due to the limited availability of fluorescent probes remaining functional after secretion. In this work, we used the Escherichia coli intimin and the Listeria monocytogenes InlB invasin as surface exposed scaffolds fused with the recently developed chemogenetic fluorescent reporter protein FAST. Using both membrane permeant (HBR-3,5DM) and non-permeant (HBRAA-3E) fluorogens that fluoresce upon binding to FAST, we demonstrated that fully functional FAST can be exposed at the cell surface and used to specifically tag the external side of the bacterial envelop in both diderm and monoderm bacteria. Our work opens new avenues to study the organization and dynamics of the bacterial cell surface proteins.Yankel ChekliCaroline Peron-CaneDario Dell’ArcipreteJean-François AllemandChenge LiJean-Marc GhigoArnaud GautierAlice LebretonNicolas DespratChristophe BeloinNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-13 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yankel Chekli Caroline Peron-Cane Dario Dell’Arciprete Jean-François Allemand Chenge Li Jean-Marc Ghigo Arnaud Gautier Alice Lebreton Nicolas Desprat Christophe Beloin Visualizing the dynamics of exported bacterial proteins with the chemogenetic fluorescent reporter FAST
description	Abstract Bacterial proteins exported to the cell surface play key cellular functions. However, despite the interest to study the localisation of surface proteins such as adhesins, transporters or hydrolases, monitoring their dynamics in live imaging remains challenging, due to the limited availability of fluorescent probes remaining functional after secretion. In this work, we used the Escherichia coli intimin and the Listeria monocytogenes InlB invasin as surface exposed scaffolds fused with the recently developed chemogenetic fluorescent reporter protein FAST. Using both membrane permeant (HBR-3,5DM) and non-permeant (HBRAA-3E) fluorogens that fluoresce upon binding to FAST, we demonstrated that fully functional FAST can be exposed at the cell surface and used to specifically tag the external side of the bacterial envelop in both diderm and monoderm bacteria. Our work opens new avenues to study the organization and dynamics of the bacterial cell surface proteins.
format	article
author	Yankel Chekli Caroline Peron-Cane Dario Dell’Arciprete Jean-François Allemand Chenge Li Jean-Marc Ghigo Arnaud Gautier Alice Lebreton Nicolas Desprat Christophe Beloin
author_facet	Yankel Chekli Caroline Peron-Cane Dario Dell’Arciprete Jean-François Allemand Chenge Li Jean-Marc Ghigo Arnaud Gautier Alice Lebreton Nicolas Desprat Christophe Beloin
author_sort	Yankel Chekli
title	Visualizing the dynamics of exported bacterial proteins with the chemogenetic fluorescent reporter FAST
title_short	Visualizing the dynamics of exported bacterial proteins with the chemogenetic fluorescent reporter FAST
title_full	Visualizing the dynamics of exported bacterial proteins with the chemogenetic fluorescent reporter FAST
title_fullStr	Visualizing the dynamics of exported bacterial proteins with the chemogenetic fluorescent reporter FAST
title_full_unstemmed	Visualizing the dynamics of exported bacterial proteins with the chemogenetic fluorescent reporter FAST
title_sort	visualizing the dynamics of exported bacterial proteins with the chemogenetic fluorescent reporter fast
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/74bfb02b768543aba2336c13542387e2
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Visualizing the dynamics of exported bacterial proteins with the chemogenetic fluorescent reporter FAST

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