RECRUITMENT OF WILD-TYPE AND RECOMBINANT ADENOASSOCIATED VIRUS INTO ADENOVIRUS REPLICATION CENTERS

Replication of a human parvovirus, adeno-associated virus (AAV), is fa cilitated by coinfection with adenovirus to provide essential helper f unctions. We have used the techniques of in situ hybridization and imm unocytochemistry to characterize the localization of AAV replication w ithin infected cells. Previous studies have shown that adenovirus esta blishes foci called replication centers within the nucleus, where aden oviral replication and transcription occur. Our studies indicate that AAV is colocalized with the adenovirus replication centers, where it m ay utilize adenovirus and cellular proteins for its own replication. E xpression of the AAV Rep protein inhibits the normal maturation of the adenovirus centers. Similar experiments were performed with recombina nt AAV (rAAV) to establish a relationship between intranuclear localiz ation and rAAV transduction. rAAV efficiently entered the cell, and it s genome was faintly detectable in a perinuclear distribution and nas mobilized to replication centers when the cell was infected with adeno virus. The recruitment of the replication-defective genome into the in tranuclear adenovirus domains resulted in enhanced transduction. These studies illustrate the importance of intracellular compartmentalizati on for such complex interactions as the relationship between AAV and a denovirus.