2HW0

NMR Solution Structure of the nuclease domain from the Replicator Initiator Protein from porcine circovirus PCV2

2HW0 の概要

• エントリーDOI: 10.2210/pdb2hw0/pdb
• 分子名称: Replicase (1 entity in total)
• 機能のキーワード: alpha+beta, hydrolase, replication
• 由来する生物種: Porcine circovirus 2
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 13275.99

構造登録者

Vega-Rocha, S.,Byeon, I.L.,Gronenborn, B.,Gronenborn, A.M.,Campos-Olivas, R. (登録日: 2006-07-31, 公開日: 2007-06-12, 最終更新日: 2024-05-29)

主引用文献

Vega-Rocha, S.,Byeon, I.L.,Gronenborn, B.,Gronenborn, A.M.,Campos-Olivas, R.
Solution structure, divalent metal and DNA binding of the endonuclease domain from the replication initiation protein from porcine circovirus 2
J.Mol.Biol., 367:473-487, 2007

PubMed Abstract: Circoviruses are the smallest circular single-stranded DNA viruses able to replicate in mammalian cells. Essential to their replication is the replication initiator, or Rep protein that initiates the rolling circle replication (RCR) of the viral genome. Here we report the NMR solution three-dimensional structure of the endonuclease domain from the Rep protein of porcine circovirus type 2 (PCV2), the causative agent of postweaning multisystemic wasting syndrome in swine. The domain comprises residues 12-112 of the full-length protein and exhibits the fold described previously for the Rep protein of the representative geminivirus tomato yellow leaf curl Sardinia virus. The structure, however, differs significantly in some secondary structure elements that decorate the central five-stranded beta-sheet, including the replacement of a beta-hairpin by an alpha-helix in PCV2 Rep. The identification of the divalent metal binding site was accomplished by following the paramagnetic broadening of NMR amide signals upon Mn(2+) titration. The site comprises three conserved acidic residues on the exposed face of the central beta-sheet. For the 1:1 complex of the PCV2 Rep nuclease domain with a 22mer double-stranded DNA oligonucleotide chemical shift mapping allowed the identification of the DNA binding site on the protein and aided in constructing a model of the protein/DNA complex.

PubMed: 17275023
DOI: 10.1016/j.jmb.2007.01.002

実験手法

SOLUTION NMR