2HOI

Crystal structure of the tetrameric pre-cleavage synaptic complex in the cre-loxp site-specific recombination

2HOI の概要

• エントリーDOI: 10.2210/pdb2hoi/pdb
• 関連するPDBエントリー: 1CRX 2HOF 4CRX
• 分子名称: LoxP DNA, Recombinase Cre, ... (4 entities in total)
• 機能のキーワード: cre-loxp precleavage synapse, recombination-dna complex, recombination/dna
• 由来する生物種: Enterobacteria phage P1
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 197186.14

構造登録者

Ghosh, K.,Van Duyne, G.D. (登録日: 2006-07-14, 公開日: 2007-06-26, 最終更新日: 2023-08-30)

主引用文献

Ghosh, K.,Guo, F.,Van Duyne, G.D.
Synapsis of loxP sites by Cre recombinase.
J.Biol.Chem., 282:24004-24016, 2007

PubMed Abstract: Cre recombinase catalyzes site-specific recombination between 34-bp loxP sites in a variety of topological and cellular contexts. An obligatory step in the recombination reaction is the association, or synapsis, of Cre-bound loxP sites to form a tetrameric protein assembly that is competent for strand exchange. Using analytical ultracentrifugation and electrophoresis approaches, we have studied the energetics of Cre-mediated synapsis of loxP sites. We found that synapsis occurs with a high affinity (Kd = 10 nM) and is pH-dependent but does not require divalent cations. Surprisingly, the catalytically inactive Cre K201A mutant is fully competent for synapsis of loxP sites, yet the inactive Y324F and R173K mutants are defective for synapsis. These findings have allowed us to determine the first crystal structures of a pre-cleavage Cre-loxP synaptic complex in a configuration representing the starting point in the recombination pathway. When combined with a quantitative analysis of synapsis using loxP mutants, the structures explain how the central 8 bp of the loxP site are able to dictate the order of strand exchange in the Cre system.

PubMed: 17573343
DOI: 10.1074/jbc.M703283200

実験手法

X-RAY DIFFRACTION (2.601 Å)