Bacteriophages are the most common biological entities and found in every biosphere. Bacteriophages can be classified into eleven families based on the nucleic acid composition, capsid symmetry, and presence or absence of a membrane or envelope. Unlike mammalian viruses, relatively few bacteriophages have an internal or exterior membrane1. Despite the fact that membrane-containing phages have been discovered for over 50 years, the structural details of the majority of the proteins still remains a mystery. How these phage proteins aid in infection cycle inside the host cell, still remains enigmatic.
In this study, we elucidated the structural intricacies of membrane-containing phages at unprecedented resolution by combining complementary methods, including biochemistry, fluorescence microscopy, and cryo-electron tomography (Cryo-ET). By employing cryo-ET and sub-tomogram averaging, we unveiled the structural details of phage spike protein, nucleocapsid and the packaging enzyme. This study offers valuable insights into the functions of phage proteins during the infection cycle within the host cell.