Anellovirus protein coded by ORF2/3 recruits host cell replication and homologous recombination machinery during replication

Anelloviridae is a family of single-stranded DNA viruses that are thought to be non-pathogenic and commensal. Despite their ubiquitous presence in human populations, little is known about the anellovirus mechanism of replication in host cells. We identified the protein coded by ORF2/3 as necessary and sufficient to initiate replication from the minimal origin of replication for viruses of both the Beta- and Alphatorquevirus genera. Supporting this observation, we identified components of the polymerase alpha and BTR complexes as interacting with the viral replication initiation protein (Rip) during DNA replication, suggesting a recombination-dependent mechanism of replication that uses host cell machinery to mediate dissolution of replication intermediates. Furthermore, we mapped a 92-bp minimal origin of replication sequence for the Betatorquevirus genus comprised of an AT-rich stretch and a portion of the GC-rich region. Altogether, this study provides a first insight into the mechanism by which anelloviruses manipulate host cell machinery to facilitate viral genome replication and represents a significant step forward in understanding the complex processes underlying anellovirus replication and persistent infection of these important commensal viruses.