Structural Insights into the Assembly of the Adeno-associated Virus Type 2 Rep68 Protein on the Integration Site AAVS1.

摘要: Adeno-associated virus (AAV) is the only eukaryotic with property of establishing latency by integrating site-specifically into human genome. The integration site known as AAVS1 located in chromosome 19 and contains multiple GCTC repeats that are recognized AAV non-structural Rep proteins. These proteins multifunctional, an N-terminal origin-binding domain (OBD) a helicase joined together short linker. As first step to understand process site-specific integration, we proceeded characterize recognition assembly Rep68 onto site. We determined x-ray structure AAV-2 OBD complex DNA Specificity achieved through interaction glycine-rich loop binds major groove α-helix interacts downstream minor on same face DNA. Although shows three molecules bound site, show using analytical centrifugation electron microscopy full-length forms heptameric complex. Moreover, minimum two direct required form stable melt Finally, although individual domains bind poorly, requires oligomerization cooperation between its OBD, helicase, linker domains.