4AAB

Crystal structure of the mutant D75N I-CreI in complex with its wild- type target (The four central bases, 2NN region, are composed by GTAC from 5' to 3')

4AAB の概要

• エントリーDOI: 10.2210/pdb4aab/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 4AAD 4AAE 4AAF 4AAG
• 分子名称: DNA ENDONUCLEASE I-CREI, 14MER DNA 5'-D(*TP*CP*AP*AP*AP*AP*CP*GP*TP*CP*GP*TP*AP*CP)-3', 10MER DNA 5'-D(*GP*AP*CP*GP*TP*TP*TP*TP*GP*AP)-3', ... (7 entities in total)
• 機能のキーワード: hydrolase-dna complex, gene targeting, protein-dna interaction, homing endonucleases, hydrolase/dna
• 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: 50154.29

構造登録者

Molina, R.,Redondo, P.,Stella, S.,Marenchino, M.,D'Abramo, M.,Gervasio, F.L.,Epinat, J.C.,Valton, J.,Grizot, S.,Duchateau, P.,Prieto, J.,Montoya, G. (登録日: 2011-12-01, 公開日: 2012-05-02, 最終更新日: 2023-12-20)

主引用文献

Molina, R.,Redondo, P.,Stella, S.,Marenchino, M.,D'Abramo, M.,Gervasio, F.L.,Charles Epinat, J.,Valton, J.,Grizot, S.,Duchateau, P.,Prieto, J.,Montoya, G.
Non-Specific Protein-DNA Interactions Control I-Crei Target Binding and Cleavage.
Nucleic Acids Res., 40:6936-6945, 2012

PubMed Abstract: Homing endonucleases represent protein scaffolds that provide powerful tools for genome manipulation, as these enzymes possess a very low frequency of DNA cleavage in eukaryotic genomes due to their high specificity. The basis of protein-DNA recognition must be understood to generate tailored enzymes that target the DNA at sites of interest. Protein-DNA interaction engineering of homing endonucleases has demonstrated the potential of these approaches to create new specific instruments to target genes for inactivation or repair. Protein-DNA interface studies have been focused mostly on specific contacts between amino acid side chains and bases to redesign the binding interface. However, it has been shown that 4 bp in the central DNA sequence of the 22-bp substrate of a homing endonuclease (I-CreI), which do not show specific protein-DNA interactions, is not devoid of content information. Here, we analyze the mechanism of target discrimination in this substrate region by the I-CreI protein, determining how it can occur independently of the specific protein-DNA interactions. Our data suggest the important role of indirect readout in this substrate region, opening the possibility for a fully rational search of new target sequences, thus improving the development of redesigned enzymes for therapeutic and biotechnological applications.

PubMed: 22495931
DOI: 10.1093/NAR/GKS320

実験手法

X-RAY DIFFRACTION (2.5 Å)