4ZQ9

X-ray structure of AAV-2 OBD bound to AAVS1 site 3:1

4ZQ9 の概要

• エントリーDOI: 10.2210/pdb4zq9/pdb
• 関連するPDBエントリー: 4ZO0
• 分子名称: Protein Rep68, DNA (5'-D(*GP*CP*GP*CP*TP*CP*GP*CP*TP*CP*GP*CP*TP*CP*GP*CP*TP*GP*GP*GP*C)-3'), DNA (5'-D(*CP*GP*CP*CP*CP*AP*GP*CP*GP*AP*GP*CP*GP*AP*GP*CP*GP*AP*GP*CP*G)-3'), ... (5 entities in total)
• 機能のキーワード: dna binding protein, adeno-associated virus, nuclease, origin-binding protein, dna binding protein-dna complex, dna binding protein/dna
• 由来する生物種: Adeno-associated virus 2 (isolate Srivastava/1982) (AAV-2); 詳細
• タンパク質・核酸の鎖数: 5
• 化学式量合計: 86240.17

構造登録者

Musayev, F.N.,Zarate-Perez, F.,Escalante, C.R. (登録日: 2015-05-08, 公開日: 2015-09-23, 最終更新日: 2024-03-06)

主引用文献

Musayev, F.N.,Zarate-Perez, F.,Bishop, C.,Burgner, J.W.,Escalante, C.R.
Structural Insights into the Assembly of the Adeno-associated Virus Type 2 Rep68 Protein on the Integration Site AAVS1.
J.Biol.Chem., 290:27487-27499, 2015

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

PubMed: 26370092
DOI: 10.1074/jbc.M115.669960

実験手法

X-RAY DIFFRACTION (2.6 Å)