PDBsum entry 7jsf

Viral protein

PDB id: 7jsf

Contents

Protein chains

(+ 1 more) 276 a.a.

DNA/RNA

PDB id:

7jsf

Name:

Viral protein

Title:

Adeno-associated virus helicase domain heptamer with ssdna

Structure:

Protein rep68. Chain: a, b, c, d, e, f, g. Engineered: yes. Mutation: yes. DNA (5'-d(p Tp Tp Tp T)-3'). Chain: h. Engineered: yes

Source:

Adeno-associated virus - 2. Aav-2. Organism_taxid: 10804. Gene: rep68. Expressed in: escherichia coli 'bl21-gold(de3)plyss ag'. Expression_system_taxid: 866768. Synthetic: yes. Synthetic construct. Organism_taxid: 32630

Authors:

C.R.Escalante

Key ref:

V.Santosh et al. (2020). The Cryo-EM structure of AAV2 Rep68 in complex with ssDNA reveals a malleable AAA+ machine that can switch between oligomeric states. Nucleic Acids Res, 48, 12983-12999. PubMed id: 33270897 DOI: 10.1093/nar/gkaa1133

Date:

14-Aug-20 Release date: 16-Dec-20

Protein chains

P03132  (REP68_AAV2S) -  Protein Rep68 from Adeno-associated virus 2 (isolate Srivastava/1982)

Seq: 536 a.a.

Struc: 276 a.a.

DNA/RNA chain

T-T-T-T 4 bases

Enzyme reactions

• Enzyme class: E.C.3.6.4.12 - Dna helicase.
• Reaction: ATP + H2O = ADP + phosphate + H⁺

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

DOI no: 10.1093/nar/gkaa1133

Nucleic Acids Res 48:12983-12999 (2020)

PubMed id: 33270897

The Cryo-EM structure of AAV2 Rep68 in complex with ssDNA reveals a malleable AAA+ machine that can switch between oligomeric states.

V.Santosh, F.N.Musayev, R.Jaiswal, F.Zárate-Pérez, B.Vandewinkel, C.Dierckx, M.Endicott, K.Sharifi, K.Dryden, E.Henckaerts, C.R.Escalante.

ABSTRACT

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.