3PLW

Ref protein from P1 bacteriophage

3PLW の概要

• エントリーDOI: 10.2210/pdb3plw/pdb
• 分子名称: Recombination enhancement function protein, ZINC ION, SULFATE ION, ... (4 entities in total)
• 機能のキーワード: hnh nuclease, dnase, hydrolase
• 由来する生物種: Enterobacteria phage P1 (Bacteriophage P1)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 21601.86

構造登録者

Keck, J.L.,Lu, D.,Cox, M.M. (登録日: 2010-11-15, 公開日: 2010-12-29, 最終更新日: 2024-02-21)

主引用文献

Gruenig, M.C.,Lu, D.,Won, S.J.,Dulberger, C.L.,Manlick, A.J.,Keck, J.L.,Cox, M.M.
Creating Directed Double-strand Breaks with the Ref Protein: A NOVEL RecA-DEPENDENT NUCLEASE FROM BACTERIOPHAGE P1.
J.Biol.Chem., 286:8240-8251, 2011

PubMed Abstract: The bacteriophage P1-encoded Ref protein enhances RecA-dependent recombination in vivo by an unknown mechanism. We demonstrate that Ref is a new type of enzyme; that is, a RecA-dependent nuclease. Ref binds to ss- and dsDNA but does not cleave any DNA substrate until RecA protein and ATP are added to form RecA nucleoprotein filaments. Ref cleaves only where RecA protein is bound. RecA functions as a co-nuclease in the Ref/RecA system. Ref nuclease activity can be limited to the targeted strands of short RecA-containing D-loops. The result is a uniquely programmable endonuclease activity, producing targeted double-strand breaks at any chosen DNA sequence in an oligonucleotide-directed fashion. We present evidence indicating that cleavage occurs in the RecA filament groove. The structure of the Ref protein has been determined to 1.4 Å resolution. The core structure, consisting of residues 77-186, consists of a central 2-stranded β-hairpin that is sandwiched between several α-helical and extended loop elements. The N-terminal 76 amino acid residues are disordered; this flexible region is required for optimal activity. The overall structure of Ref, including several putative active site histidine residues, defines a new subclass of HNH-family nucleases. We propose that enhancement of recombination by Ref reflects the introduction of directed, recombinogenic double-strand breaks.

PubMed: 21193392
DOI: 10.1074/jbc.M110.205088

実験手法

X-RAY DIFFRACTION (1.4 Å)