4AQX

Crystal structure of I-CreI complexed with its target methylated at position plus 2 (in the b strand) in the presence of magnesium

4AQX の概要

• エントリーDOI: 10.2210/pdb4aqx/pdb
• 関連するPDBエントリー: 1AF5 1BP7 1G9Y 1G9Z 1MOW 1N3E 1N3F 1T9I 1T9J 1U0C 1U0D 2VBJ 2VBL 2VBN 2VBO 4AAB 4AAD 4AAE 4AAF 4AAG 4AQU
• 分子名称: DNA ENDONUCLEASE I-CREI, 5'-D(*TP*CP*AP*AP*AP*AP*CP*GP*TP*CP*GP*TP*GP*AP)-3', 5'-D(*GP*AP*CP*AP*GP*TP*TP*TP*GP*GP)-3', ... (8 entities in total)
• 機能のキーワード: hydrolase, methylation, gene targeting, genetics, protein-dna interaction, homing endonucleases
• 由来する生物種: CHLAMYDOMONAS REINHARDTII; 詳細
• 細胞内の位置: Plastid, chloroplast: P05725
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 50228.28

構造登録者

Valton, J.,Daboussi, F.,Leduc, S.,Redondo, P.,Macmaster, R.,Molina, R.,Montoya, G.,Duchateau, P. (登録日: 2012-04-19, 公開日: 2012-07-04, 最終更新日: 2023-12-20)

主引用文献

Valton, J.,Daboussi, F.,Leduc, S.,Molina, R.,Redondo, P.,Macmaster, R.,Montoya, G.,Duchateau, P.
5'-Cytosine-Phosphoguanine (Cpg) Methylation Impacts the Activity of Natural and Engineered Meganucleases.
J.Biol.Chem., 287:30139-, 2012

PubMed Abstract: In this study, we asked whether CpG methylation could influence the DNA binding affinity and activity of meganucleases used for genome engineering applications. A combination of biochemical and structural approaches enabled us to demonstrate that CpG methylation decreases I-CreI DNA binding affinity and inhibits its endonuclease activity in vitro. This inhibition depends on the position of the methylated cytosine within the DNA target and was almost total when it is located inside the central tetrabase. Crystal structures of I-CreI bound to methylated cognate target DNA suggested a molecular basis for such inhibition, although the precise mechanism still has to be specified. Finally, we demonstrated that the efficacy of engineered meganucleases can be diminished by CpG methylation of the targeted endogenous site, and we proposed a rational design of the meganuclease DNA binding domain to alleviate such an effect. We conclude that although activity and sequence specificity of engineered meganucleases are crucial parameters, target DNA epigenetic modifications need to be considered for successful gene editions.

PubMed: 22740697
DOI: 10.1074/JBC.M112.379966

実験手法

X-RAY DIFFRACTION (2.2 Å)