The Cryo-EM structure of AAV2 Rep68 in complex with ssDNA reveals a malleable AAA+ machine that can switch between oligomeric states
| Autor: | Kamyar Sharifi, Vishaka Santosh, Francisco Zarate-Perez, Els Henckaerts, Kelly A. Dryden, Carlos R. Escalante, Molly Endicott, Rahul Jaiswal, Bram Vandewinkel, Caroline Dierckx, Faik N. Musayev |
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| Rok vydání: | 2020 |
| Předmět: |
Conformational change
AcademicSubjects/SCI00010 Cryo-electron microscopy viruses Protein subunit DNA Single-Stranded Biology medicine.disease_cause Viral Proteins chemistry.chemical_compound Adenosine Triphosphate Structural Biology Genetics medicine Humans Adeno-associated virus Cryoelectron Microscopy DNA Helicases DNA replication Helicase Dependovirus DNA-Binding Proteins chemistry Biophysics biology.protein DNA Binding domain |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gkaa1133 |
| Popis: | The adeno-associated virus (AAV) non-structural Rep proteins catalyze all the DNA transactions required for virus viability including, DNA replication, transcription regulation, genome packaging, and during the latent phase, site-specific integration. Rep proteins contain two multifunctional domains: an Origin Binding Domain (OBD) and a SF3 helicase domain (HD). Studies have shown that Rep proteins have a dynamic oligomeric behavior where the nature of the DNA substrate molecule modulates its oligomeric state. In the presence of ssDNA, Rep68 forms a large double-octameric ring complex. To understand the mechanisms underlying AAV Rep function, we investigated the cryo-EM and X-ray structures of Rep68-ssDNA complexes. Surprisingly, Rep68 generates hybrid ring structures where the OBD forms octameric rings while the HD forms heptamers. Moreover, the binding to ATPγS promotes a large conformational change in the entire AAA+ domain that leads the HD to form both heptamer and hexamers. The HD oligomerization is driven by an interdomain linker region that acts as a latch to 'catch' the neighboring HD subunit and is flexible enough to permit the formation of different stoichiometric ring structures. Overall, our studies show the structural basis of AAV Rep's structural flexibility required to fulfill its multifunctional role during the AAV life cycle. ispartof: NUCLEIC ACIDS RESEARCH vol:48 issue:22 pages:12983-12999 ispartof: location:England status: published |
| Databáze: | OpenAIRE |
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