Filamentous phage infection: Required interactions with the TolA protein

Infection of Escherichia coli by the filamentous phage fl is initiated by binding of the phage to the tip of the F conjugative pilus via the gene III protein, Subsequent translocation of phage DNA requires the chromosomally encoded TolQ, TolR, and TolA proteins, after the pilus presumably has withdrawn, bringing the phage to the bacterial surface. Of these three proteins, TolA is proposed to span the periplasm, since it contains a long helical domain (domain II), which connects a cytoplasmic membrane anchor domain (domain I) to the carboxyl-terminal domain (domain III), By using a transducing phage, the requirement for TolA in an F+ strain was found to be absolute. The role of TolA domains II and III in the infective process was examined by analyzing the ability of various deletion mutants of tolA to facilitate infection, The C-terminal domain III was shown to be essential, whereas the polyglycine region separating domains I and II could be deleted with no effect, Deletion of helical domain II. reduced the efficiency of infection, which could be restored to normal by retaining the C-terminal half of domain II, Soluble domain III, expressed in the periplasm but not in the cytoplasm or in the medium, interfered with infection of a tolA(+) strain, The essential interaction of TolA domain III with phage via gene III protein appears to require interaction with a third component, either the pilus tip or a periplasmic entity.