5U91

Crystal structure of Tre/loxLTR complex

5U91 の概要

• エントリーDOI: 10.2210/pdb5u91/pdb
• 分子名称: Tre recombinase protein, DNA (37-MER), ... (4 entities in total)
• 機能のキーワード: cre mutant tre recombinase, isomerase-dna complex, isomerase/dna
• 由来する生物種: synthetic construct; 詳細
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 200710.94

構造登録者

Meinke, G.,Karpinski, J.,Buchholz, F.,Bohm, A. (登録日: 2016-12-15, 公開日: 2017-07-26, 最終更新日: 2024-11-06)

主引用文献

Meinke, G.,Karpinski, J.,Buchholz, F.,Bohm, A.
Crystal structure of an engineered, HIV-specific recombinase for removal of integrated proviral DNA.
Nucleic Acids Res., 45:9726-9740, 2017

PubMed Abstract: As part of the HIV infection cycle, viral DNA inserts into the genome of host cells such that the integrated DNA encoding the viral proteins is flanked by long terminal repeat (LTR) regions from the retrovirus. In an effort to develop novel genome editing techniques that safely excise HIV provirus from cells, Tre, an engineered version of Cre recombinase, was designed to target a 34-bp sequence within the HIV-1 LTR (loxLTR). The sequence targeted by Tre lacks the symmetry present in loxP, the natural DNA substrate for Cre. We report here the crystal structure of a catalytically inactive (Y324F) mutant of this engineered Tre recombinase in complex with the loxLTR DNA substrate. We also report that 17 of the 19 amino acid changes relative to Cre contribute to the altered specificity, even though many of these residues do not contact the DNA directly. We hypothesize that some mutations increase the flexibility of the Cre tetramer and that this, along with flexibility in the DNA, enable the engineered enzyme and DNA substrate to adopt complementary conformations.

PubMed: 28934476
DOI: 10.1093/nar/gkx603

実験手法

X-RAY DIFFRACTION (3.104 Å)