Bacteriophage P22 SieA-mediated superinfection exclusion.
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A single chromosomal copy of P22 sieA is sufficient to lower P22 plaque formation by at least seven orders of magnitude, with no effect on host growth.
"Thus, the sieA gene in one copy per Salmonella genome is sufficient to mediate robust superinfection exclusion."
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SieA blocks DNA injection itself: infection of a sieA+ strain causes almost no K+ efflux, whereas the isogenic sieA- strain releases nearly all cellular K+.
"Figure 2A shows that P22 causes very little, if any, K+ release from the sieA+ strain, while nearly all of the cellular K+ is released from the isogenic sieA– strain, suggesting that DNA injection per se is blocked by SieA."
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FLAG-tagged, exclusion-competent SieA fractionates with the inner membrane and is absent from the outer membrane.
"We conclude that SieA is an exclusively inner membrane protein."
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Escape from SieA requires amino acid changes in the C-terminal regions of the ejection proteins gp16 and gp20, implicating the DNA delivery conduit as the target.
"Since amino acid changes in E-proteins gp16 and gp20 confer escape from SieA exclusion, we conclude that SieA very likely blocks P22 DNA injection by interfering with the functions of these two proteins and that the mutations avoid this interference."
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Exclusion is specific for P22-like phages; unrelated tailed phages, including the short-tailed phage SP6, plate equally on sieA+ and sieA- hosts.
"Since SP6 is not affected, the P22 SieA protein does not exclude all short-tailed phages and appears to be specific for P22-like phages."