Bacteriophage P22 SieA mediated superinfection exclusion

Abstract

ABSTRACTMany temperate phages encode prophage-expressed functions that interfere with superinfection of the host bacterium by external phages.Salmonellaphage P22 has four such systems that are expressed from the prophage in a lysogen that are encoded by thec2(repressor),gtrABC,sieA, andsieBgenes. Here we report that the P22-encoded SieA protein is the only phage protein required for exclusion by the SieA system, and that it is an inner membrane protein that blocks DNA injection by P22 and its relatives, but has no effect on infection by other tailed phage types. The P22 virion injects its DNA through the host cell membranes and periplasm via a conduit assembled from three “ejection proteins” after their release from the virion. Phage P22 mutants were isolated that overcome the SieA block, and they have amino acid changes in the C-terminal regions of the gene16and20encoded ejection proteins. Three different single amino acid changes in these proteins are required to obtain nearly full resistance to SieA. Hybrid P22 phages that have phage HK620 ejection protein genes are also partially resistant to SieA. There are three sequence types of extant phage-encoded SieA proteins that are less than 30% identical to one another, yet comparison of two of these types found no differences in target specificity. Our data are consistent with a model in which the inner membrane protein SieA interferes with the assembly or function of the periplasmic gp20 and membrane-bound gp16 DNA delivery conduit.HIGHLIGHTSPhage P22 SieA protein blocks DNA injection by P22-like phagesSieA is an inner membrane proteinHybrid P22 phages with phage HK620 ejection proteins partially escape SieA exclusionSieA escape mutants of P22 alter the gp16 and 20 proteins that form the DNA ejection tube.