4KTW

Lactococcus phage 67 RuvC - apo form

4KTW の概要

• エントリーDOI: 10.2210/pdb4ktw/pdb
• 関連するPDBエントリー: 4KTZ
• 分子名称: RuvC endonuclease, 1,2-ETHANEDIOL, GLYCEROL, ... (4 entities in total)
• 機能のキーワード: endonuclease, dna junctions, replication, recombination, phage packaging, holliday junction, rnase-h fold, dna junction endonuclease, holliday junctions and other 2- & 3-way dna junctions, hydrolase
• 由来する生物種: Lactococcus phage bIL67
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 36399.40

構造登録者

Rafferty, J.B.,Green, V. (登録日: 2013-05-21, 公開日: 2013-08-14, 最終更新日: 2024-02-28)

主引用文献

Green, V.,Curtis, F.A.,Sedelnikova, S.,Rafferty, J.B.,Sharples, G.J.
Mutants of phage bIL67 RuvC with enhanced Holliday junction binding selectivity and resolution symmetry.
Mol.Microbiol., 89:1240-1258, 2013

PubMed Abstract: Viral and bacterial Holliday junction resolvases differ in specificity with the former typically being more promiscuous, acting on a variety of branched DNA substrates, while the latter exclusively targets Holliday junctions. We have determined the crystal structure of a RuvC resolvase from bacteriophage bIL67 to help identify features responsible for DNA branch discrimination. Comparisons between phage and bacterial RuvC structures revealed significant differences in the number and position of positively-charged residues in the outer sides of the junction binding cleft. Substitutions were generated in phage RuvC residues implicated in branch recognition and six were found to confer defects in Holliday junction and replication fork cleavage in vivo. Two mutants, R121A and R124A that flank the DNA binding site were purified and exhibited reduced in vitro binding to fork and linear duplex substrates relative to the wild-type, while retaining the ability to bind X junctions. Crucially, these two variants cleaved Holliday junctions with enhanced specificity and symmetry, a feature more akin to cellular RuvC resolvases. Thus, additional positive charges in the phage RuvC binding site apparently stabilize productive interactions with branched structures other than the canonical Holliday junction, a feature advantageous for viral DNA processing but deleterious for their cellular counterparts.

PubMed: 23888987
DOI: 10.1111/mmi.12343

実験手法

X-RAY DIFFRACTION (1.68 Å)