The mechanism of cytoplasmic orthopoxvirus DNA replication

Orthopoxvirus DNA replication occurs in the cytoplasm of infected cells within discrete foci designated as virosomes. We show that newly synthesized rabbit poxvirus (RPV) virosomal DNA consists predominantly of concatamers wherein unit length molecules are joined by fusion of two left (LL) or right (RR) ends, resulting in genomes aligned in alternating head-to-head and tail-to tail mirror image arrays. These concatameric molecules serve as the substrates from which unit length DNA molecules are excised during morphogenesis. We propose a mechanism by which internal deletions within these concatameric arrays prior to genome excision and packaging could create inverted terminal repeats and generate gene duplications.