A Chimeric Protein Containing the N Terminus of the Adeno-Associated Virus Rep Protein Recognizes Its Target Site in an In Vivo Assay

摘要: The Rep78 and Rep68 proteins of adeno-associated virus (AAV) type 2 are involved in DNA replication, regulation gene expression, targeting site-specific integration. They bind to a specific Rep recognition sequence (RRS) found both the viral inverted terminal repeats AAVS1 integration locus on human chromosome 19. Previous vitro studies implied that an N-terminal segment is recognition, while additional domains might stabilize binding mediate multimerization. In order define minimal requirements for recognize its target site genome, we developed one-hybrid assays which DNA-protein interactions detected vivo. Chimeric consisting N terminus fused different oligomerization motifs transcriptional activation domain were analyzed oligomerization, binding, reporter expression. Expression genes was driven from RRS cloned upstream promoters examined mammalian cells transfected plasmids Saccharomyces cerevisiae cassette integrated into yeast genome. Our results show first time chimeric containing amino-terminal 244 residues able vivo when incorporated artificial multimers. These suggest may be used harness unique feature AAV therapy applications.