5DCX
Structural studies of AAV2 Rep68 reveal a partially structured linker and compact domain conformation
Summary for 5DCX
| Entry DOI | 10.2210/pdb5dcx/pdb |
| Descriptor | Protein Rep68, MAGNESIUM ION (3 entities in total) |
| Functional Keywords | parvovirus, adeno-associated virus, dna replication, small-angle x-ray scattering, dna binding protein, rep proteins |
| Biological source | Adeno-associated virus 2 (AAV-2) |
| Total number of polymer chains | 11 |
| Total formula weight | 287406.53 |
| Authors | Musayev, F.N.,Zarate-Perez, F. (deposition date: 2015-08-24, release date: 2015-09-23, Last modification date: 2023-09-27) |
| Primary citation | Musayev, F.N.,Zarate-Perez, F.,Bardelli, M.,Bishop, C.,Saniev, E.F.,Linden, R.M.,Henckaerts, E.,Escalante, C.R. Structural Studies of AAV2 Rep68 Reveal a Partially Structured Linker and Compact Domain Conformation. Biochemistry, 54:5907-5919, 2015 Cited by PubMed Abstract: Adeno-associated virus (AAV) nonstructural proteins Rep78 and Rep68 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 SF3 helicase domain. A short linker of ∼20 amino acids that is critical for oligomerization and function connects the two domains. Although X-ray structures of the AAV5 OBD and AAV2 helicase domains have been determined, information about the full-length protein and linker conformation is not known. This article presents the solution structure of AAV2 Rep68 using small-angle X-ray scattering (SAXS). We first determined the X-ray structures of the minimal AAV2 Rep68 OBD and of the OBD with the linker region. These X-ray structures reveal novel features that include a long C-terminal α-helix that protrudes from the core of the protein at a 45° angle and a partially structured linker. SAXS studies corroborate that the linker is not extended, and we show that a proline residue in the linker is critical for Rep68 oligomerization and function. SAXS-based rigid-body modeling of Rep68 confirms these observations, showing a compact arrangement of the two domains in which they acquire a conformation that positions key residues in all domains on one face of the protein, poised to interact with DNA. PubMed: 26314310DOI: 10.1021/acs.biochem.5b00610 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.6 Å) |
Structure validation
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