CryoEM Studies of AAV Rep-DNA complexes underlying AAV life cycle.
EMSL Project ID
50754
Abstract
Adeno-associated virus (AAV) non-structural Rep proteins carry out all DNA transactions that regulate the AAV life cycle. They share two multifunctional domains: an N-terminal origin binding/nicking domain (OBD) from the HUH superfamily and a C-terminal SF3 helicase domain. The two domains are linked by a short region of ~ 20 amino acids that is critical for oligomerization. The two domains have multiple DNA interfaces that allow Rep proteins to form Rep-DNA complexes with different stoichiometries to fulfill specific functions. We have determined that in the presence of ssDNA poly-dT, Rep68 forms double octameric rings arranged in a head to head orientation. In contrast, Rep68 forms an heptameric complex when bound to the integration site AAVS1. In this proposal, we will determine the structural landscape of how Rep proteins form different complexes with DNA, each fine-tuned for a specific process including DNA replication, regulation of transcription, site-specific integration and genome packaging.
Project Details
Start Date
2019-04-15
End Date
2021-03-17
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members