2I3P

K28R mutant of Homing Endonuclease I-CreI

2I3P の概要

• エントリーDOI: 10.2210/pdb2i3p/pdb
• 関連するPDBエントリー: 2I3Q
• 分子名称: 5'-D(*GP*CP*AP*AP*AP*TP*CP*GP*TP*CP*GP*TP*GP*AP*GP*AP*CP*AP*AP*TP*TP*TP*CP*G)-3', 5'-D(*CP*GP*AP*AP*AP*TP*TP*GP*TP*CP*TP*CP*AP*CP*GP*AP*CP*GP*AP*TP*TP*TP*GP*C)-3', DNA endonuclease I-CreI, ... (5 entities in total)
• 機能のキーワード: homing endonulease i-crei, dna, hydrolase-dna complex, hydrolase/dna
• 由来する生物種: Chlamydomonas reinhardtii
• 細胞内の位置: Plastid, chloroplast: P05725
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 50169.36

構造登録者

Sussman, D.,Rosen, L. (登録日: 2006-08-20, 公開日: 2006-09-05, 最終更新日: 2023-08-30)

主引用文献

Rosen, L.E.,Morrison, H.A.,Masri, S.,Brown, M.J.,Springstubb, B.,Sussman, D.,Stoddard, B.L.,Seligman, L.M.
Homing endonuclease I-CreI derivatives with novel DNA target specificities.
Nucleic Acids Res., 34:4791-4800, 2006

PubMed Abstract: Homing endonucleases are highly specific enzymes, capable of recognizing and cleaving unique DNA sequences in complex genomes. Since such DNA cleavage events can result in targeted allele-inactivation and/or allele-replacement in vivo, the ability to engineer homing endonucleases matched to specific DNA sequences of interest would enable powerful and precise genome manipulations. We have taken a step-wise genetic approach in analyzing individual homing endonuclease I-CreI protein/DNA contacts, and describe here novel interactions at four distinct target site positions. Crystal structures of two mutant endonucleases reveal the molecular interactions responsible for their altered DNA target specificities. We also combine novel contacts to create an endonuclease with the predicted target specificity. These studies provide important insights into engineering homing endonucleases with novel target specificities, as well as into the evolution of DNA recognition by this fascinating family of proteins.

PubMed: 16971456
DOI: 10.1093/nar/gkl645

実験手法

X-RAY DIFFRACTION (2.3 Å)