Deciphering the assembly of multimolecular membrane protein complexes in bacteria using two-hybrid assays
Conférence organisée par le laboratoire SABNP.
Par Daniel Ladant - Institut Pasteur
Unité de Biochimie des Interactions Macromoléculaires, CNRS UMR 3528
Département de Biologie Structurale et Chimie
Le 17 avril à 14h
Bâtiment Maupertuis - salle 02 W34
3 rue du Père Jarlan - Evry
Protein-protein interactions (PPIs) and multi-protein complexes perform central roles in all living organisms. Genetic assays capable of detecting protein interactions within living, intact cells have been particularly useful to identify and characterize protein interaction networks. We previously designed a bacterial two-hybrid system, BACTH (Bacterial Adenylate Cyclase-based Two-Hybrid), that is based on the interaction-mediated reconstitution of a cAMP signaling cascade in Escherichia coli. This technique is particularly appropriate to characterize interactions between membrane-associated proteins. I will describe our study of the assembly of the bacterial cell division machinery or divisiome, a membrane-associated, multimolecular complex made of more than a dozen of essential proteins (called Fts) and that assemble at the future cell division site to trigger the septation of a bacterium into two daughter cells. I will also present further implementations of the BACTH system that greatly increase the sensitivity of this genetic system to reach single-molecule sensitivity.