TY - JOUR T1 - Visualizing the dynamics of exported bacterial proteins with the chemogenetic fluorescent reporter FAST JF - bioRxiv DO - 10.1101/2020.05.12.090142 SP - 2020.05.12.090142 AU - Yankel Chekli AU - Caroline Peron-Cane AU - Dario Dell’Arciprete AU - Jean-François Allemand AU - Chenge Li AU - Jean-Marc Ghigo AU - Arnaud Gautier AU - Alice Lebreton AU - Nicolas Desprat AU - Christophe Beloin Y1 - 2020/01/01 UR - http://biorxiv.org/content/early/2020/05/13/2020.05.12.090142.1.abstract N2 - 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. ER -