5AKF

THE CRYSTAL STRUCTURE OF I-DMOI Q42AK120M IN COMPLEX WITH ITS TARGET DNA NICKED IN THE CODING STRAND A AND IN THE PRESENCE OF 2MM MN

5AKF の概要

• エントリーDOI: 10.2210/pdb5akf/pdb
• 関連するPDBエントリー: 5AK9 5AKM 5AKN
• 分子名称: HOMING ENDONUCLEASE I-DMOI, 5'-D(*GP*CP*CP*TP*TP*GP*CP*CP*GP*GP*GP*TP*AP*AP)-3', 5'-D(*GP*TP*TP*CP*CP*GP*GP*CP*GP*CP*GP)-3', ... (8 entities in total)
• 機能のキーワード: hydrolase, gene targeting, genetics, protein-dna interaction, homing endonucleases
• 由来する生物種: DESULFUROCOCCUS MOBILIS; 詳細
• タンパク質・核酸の鎖数: 12
• 化学式量合計: 115949.44

構造登録者

Molina, R.,Marcaida, M.J.,Redondo, P.,Marenchino, M.,D'Abramo, M.,Montoya, G.,Prieto, J. (登録日: 2015-03-03, 公開日: 2015-06-17, 最終更新日: 2024-01-10)

主引用文献

Molina, R.,Marcaida, M.J.,Redondo, P.,Marenchino, M.,Duchateau, P.,D'Abramo, M.,Montoya, G.,Prieto, J.
Engineering a Nickase on the Homing Endonuclease I-Dmoi Scaffold.
J.Biol.Chem., 290:18534-, 2015

PubMed Abstract: Homing endonucleases are useful tools for genome modification because of their capability to recognize and cleave specifically large DNA targets. These endonucleases generate a DNA double strand break that can be repaired by the DNA damage response machinery. The break can be repaired by homologous recombination, an error-free mechanism, or by non-homologous end joining, a process susceptible to introducing errors in the repaired sequence. The type of DNA cleavage might alter the balance between these two alternatives. The use of "nickases" producing a specific single strand break instead of a double strand break could be an approach to reduce the toxicity associated with non-homologous end joining by promoting the use of homologous recombination to repair the cleavage of a single DNA break. Taking advantage of the sequential DNA cleavage mechanism of I-DmoI LAGLIDADG homing endonuclease, we have developed a new variant that is able to cut preferentially the coding DNA strand, generating a nicked DNA target. Our structural and biochemical analysis shows that by decoupling the action of the catalytic residues acting on each strand we can inhibit one of them while keeping the other functional.

PubMed: 26045557
DOI: 10.1074/JBC.M115.658666

実験手法

X-RAY DIFFRACTION (2.45 Å)