Structural Dynamics of Bacteriophage P22 Virions During the Initiation of Infection

For successful infection, bacteriophages must overcome multiple barriers to transport the genome and proteins across the bacterial cell envelope. We use cryo-electron tomography to study infection initiation of phage P22 in Salmonella enterica sv. Typhimurium, revealing how a channel forms to allow genome translocation into the cytoplasm. Our results show free phages initially attaching obliquely to the cell through interactions between the O antigen and two of the six tailspikes; the tail needle also abuts the cell surface. The virion then orients to the perpendicular and the needle penetrates the outer membrane. The needle is released and the internal head protein gp7* is ejected and assembles into an extra-cellular channel extending from the gp10 baseplate to the cell surface. A second protein, gp20, is ejected and assembles into a structure that extends the extra-cellular channel across the outer membrane into the periplasm. Insertion of the third ejected protein gp16 into the cytoplasmic membrane likely completes the overall trans-envelope channel into the cytoplasm. Construction of a trans-envelope channel is an essential step during infection by all short-tailed phages of Gram-negative bacteria because such virions cannot directly deliver their genome into the cell cytoplasm.