RT Journal Article
SR Electronic
T1 Visualizing the dynamics of exported bacterial proteins with the chemogenetic fluorescent reporter FAST
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.05.12.090142
DO 10.1101/2020.05.12.090142
A1 Yankel Chekli
A1 Caroline Peron-Cane
A1 Dario Dell’Arciprete
A1 Jean-François Allemand
A1 Chenge Li
A1 Jean-Marc Ghigo
A1 Arnaud Gautier
A1 Alice Lebreton
A1 Nicolas Desprat
A1 Christophe Beloin
YR 2020
UL http://biorxiv.org/content/early/2020/05/13/2020.05.12.090142.1.abstract
AB Bacterial proteins exported to the cell surface play key cellular functions. However, despite the interest to study the localization 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 specifically tagged on the external side of the bacterial envelop in both diderm and monoderm bacteria. Our work opens new avenues to study of the organization and dynamics of the bacterial cell surface proteins.Competing Interest StatementThe authors have declared no competing interest.