Abstract
Despite tight genetic compression, viral genomes are often organized into functional gene clusters, a modular structure that might favor their evolvability. This has greatly facilitated biotechnological developments such as the recombinant adeno-associated virus (AAV) systems for gene therapy. Following this lead, we endeavored to engineer the related insect parvovirus Junonia coenia densovirus (JcDV) to create addressable vectors for insect pest biocontrol. To enable safer manipulation of capsid mutants, we translocated the nonstructural (ns) gene cluster outside the viral genome. To our dismay, this yielded a virtually nonreplicable clone. We linked the replication defect to an unexpected modularity breach, as ns translocation truncated the overlapping 3' untranslated region (UTR) of the capsid transcript (vp). We found that the native vp 3' UTR is necessary for high-level VP production but that decreased expression does not adversely impact the expression of NS proteins, which are known replication effectors. As nonsense vp mutations recapitulate the replication defect, VP proteins appear to be directly implicated in the replication process. Our findings suggest intricate replication-encapsidation couplings that favor the maintenance of genetic integrity. We discuss possible connections with an intriguing cis-packaging phenomenon previously observed in parvoviruses whereby capsids preferentially package the genome from which they were expressed.