A short noncoding viral DNA element showing characteristics of a replication origin confers bacteriophage resistance to Streptococcus thermophilus.
A 302-bp noncoding DNA fragment from the DNA replication module of phage phiSfi21 was shown to protect the Streptococcus thermophilus strain Sfi1 from infection by 17 of 25 phages. The phage-inhibitory DNA possesses two determinants, each of which individually mediated phage resistance. The phage-inhibitory activity was copy number dependent and operates by blocking the accumulation of phage DNA. Furthermore, when cloned on a plasmid, the phiSfi21 DNA acts as an origin of replication driven by phage infection. Protein or proteins in the phiSfi21-infected cells were shown to interact with this phage-inhibitory DNA fragment, forming a retarded protein-DNA complex in gel retardation assays. A model in which phage proteins interact with the inhibitory DNA such that they are no longer available for phage propagation can be used to explain the observed bacteriophage resistance. Genome analysis of phiSfi19, a phage that is insensitive to the inhibitory activity of the phiSfi21-derived DNA, led to the characterisation of a variant putative phage replication origin that differed in 14 of 302 nucleotides from that of phiSfi21. The variant origin was cloned and exhibited an inhibitory activity toward phages that were insensitive to the phiSfi21-derived DNA.
Foley
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Nestlé Research Centre, Nestec Ltd., Vers-chez-les-Blanc, CH-1000, Lausanne, 26, Switzerland.
Lucchini
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Zwahlen
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MC
Brüssow
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D001483:Base Sequence; D004261:DNA Replication; D004279:DNA, Viral; D018628:Gene Dosage; D008969:Molecular Sequence Data; D010641:Phenotype; D010957:Plasmids; D018741:Replication Origin; D013291:Streptococcus / Q000821:virology; D017108:Streptococcus Phages / Q000235:genetics* / Q000502:physiology; D014779:Virus Replication
D004279:DNA, Viral