Cryo-electron tomography study of bacterial wall perforation by bacteriophage T5

The project involves obtaining the structure of the T5 channel perforating the bacterial wall by in vivo cryo-electron tomography, to understand how the phage manages to cross the outer membrane and then the inner membrane of a minicell, and to identify and understand the role of host factors. Minicells result from an abnormal division of E. coli, leading to the formation of small, round, nucleoid-free cells of a diameter compatible with electron microscopy imaging. At phage level, we have already determined, at atomic resolution, the structure of the tail end of T5 before and after interaction with FhuA reconstituted in nanodiscs: this latter structure will be invaluable for fitting it into the sub-tomogram of averaged structures obtained by cryo-electron tomography, to understand at atomic level the perforation of the E. coli cell wall.