Abstract
Many DNA viruses concatemerize their genomes as a prerequisite to packaging into capsids. Concatemerization arises from either replication or homologous recombination. Replication is already the target of many antiviral drugs, and viral recombinases are an attractive target for drug design, particularly for combination therapy with replication inhibitors, due to their important supporting role in viral growth. To dissect the molecular mechanisms of viral recombination, we and others previously identified a family of viral nucleases that comprise one component of a conserved, two-component viral recombination system. The nuclease component is related to the exonuclease of phage λ and is common to viruses with linear double-stranded DNA genomes. To test the idea that these viruses have a common strategy for recombination and genome concatemerization, we isolated the previously uncharacterized
34.1
gene from
Bacillus subtilis
phage SPP1, expressed it in
Escherichia coli
, purified the protein, and determined its enzymatic properties. Like λ exonuclease, Chu (the product of
34.1
) forms an oligomer, is a processive alkaline exonuclease that digests linear double-stranded DNA in a Mg
2+
-dependent reaction, and shows a preference for 5′-phosphorylated DNA ends. A model for viral recombination, based on the phage λ Red recombination system, is proposed.
