Bacteriophages recognize and bind specific receptors to infect suitable hosts. Bacteriophage SPP1 targets at least two receptors of the Bacillus subtilis cell envelope, the glucosylated wall teichoic acids and the membrane protein YueB. Here, we identify a key virion protein for YueB binding and for the trigger of DNA ejection. Extracts from B. subtilis-infected cells applied to a YueB affinity matrix led to preferential capturing of gp21 from SPP1. To assess the significance of this interaction, we isolated mutant phages specifically affected in YueB binding. The mutants exhibited a very low inactivation rate and a strong defect to eject DNA when challenged with YueB. The phenotype correlated with presence of a single amino acid substitution in the gp21 carboxyl terminus, defining a region involved in YueB binding. Immunoelectron microscopy located the gp21 N-terminus in the SPP1 cap and probably in the adjacent tail spike region whereas the gp21 C-terminus was mapped further down in the spike structure. Antibodies against this part of gp21 interfered with the interaction of YueB with SPP1 and triggered DNA ejection. The gp21 C-terminal region thus plays a central role in two early key events that commit the virus to deliver its genome into host cells.
© 2011 Blackwell Publishing Ltd.