Rescue and Autonomous Replication of Adeno-Associated Virus Type 2 Genomes Containing Rep-Binding Site Mutations in the Viral p5 Promoter

摘要: The Rep proteins encoded by the adeno-associated virus type 2 (AAV) play a crucial role in rescue, replication, and integration of viral genome. In absence helper virus, little expression AAV occurs, genome fails to undergo DNA replication. Since previous studies have established that Rep78 Rep68 from p5 promoter is controlled Rep-binding site (RBS) YY1 factor-binding (YBS), we constructed number recombinant plasmids containing mutations and/or deletions RBS YBS promoter. These were transfected HeLa or 293 cells analyzed for potential rescue Our revealed (i) low-level autonomous replication wild-type occurred but not cells; (ii) resulted augmented promoter, leading more efficient (iii) alone coinfection with adenovirus; (iv) adenovirus E1A gene products was insufficient mediate (v) autonomously replicated genomes successfully encapsidated mature progeny virions biologically active secondary infection presence (vi) stable transfection resistance neomycin significantly affected only cells, presumably because these KB cells. data suggest adenovirus, protein-RBS interaction plays dominant down-regulating perturbation this sufficient confer competence