2VS8

The crystal structure of I-DmoI in complex with DNA and Mn

Summary for 2VS8

• Entry DOI: 10.2210/pdb2vs8/pdb
• Related: 1B24 1MOW 2VS7 2VS9
• Descriptor: HOMING ENDONUCLEASE I-DMOI, 5'-D(*GP*CP*CP*TP*TP*GP*CP*CP*GP*GP *GP*TP*AP*A)-3', 5'-D(*GP*TP*TP*CP*CP*GP*GP*CP* DGP*DCP*DGP)-3', ... (8 entities in total)
• Functional Keywords: meganuclease, intron homing, genetic engineering, homing endonuclease, protein/dna crystallography, nuclease, hydrolase, magnesium, endonuclease, dna-binding protein, dna binding protein
• Biological source: DESULFUROCOCCUS MOBILIS; More
• Total number of polymer chains: 15
• Total formula weight: 116514.35

Authors

Marcaida, M.J.,Prieto, J.,Redondo, P.,Nadra, A.D.,Alibes, A.,Serrano, L.,Grizot, S.,Duchateau, P.,Paques, F.,Blanco, F.J.,Montoya, G. (deposition date: 2008-04-21, release date: 2008-11-11, Last modification date: 2023-12-13)

Primary citation

Marcaida, M.J.,Prieto, J.,Redondo, P.,Nadra, A.D.,Alibes, A.,Serrano, L.,Grizot, S.,Duchateau, P.,Paques, F.,Blanco, F.J.,Montoya, G.
Crystal Structure of I-Dmoi in Complex with its Target DNA Provides New Insights Into Meganuclease Engineering.
Proc.Natl.Acad.Sci.USA, 105:16888-, 2008

PubMed Abstract: Homing endonucleases, also known as meganucleases, are sequence-specific enzymes with large DNA recognition sites. These enzymes can be used to induce efficient homologous gene targeting in cells and plants, opening perspectives for genome engineering with applications in a wide series of fields, ranging from biotechnology to gene therapy. Here, we report the crystal structures at 2.0 and 2.1 A resolution of the I-DmoI meganuclease in complex with its substrate DNA before and after cleavage, providing snapshots of the catalytic process. Our study suggests that I-DmoI requires only 2 cations instead of 3 for DNA cleavage. The structure sheds light onto the basis of DNA binding, indicating key residues responsible for nonpalindromic target DNA recognition. In silico and in vivo analysis of the I-DmoI DNA cleavage specificity suggests that despite the relatively few protein-base contacts, I-DmoI is highly specific when compared with other meganucleases. Our data open the door toward the generation of custom endonucleases for targeted genome engineering using the monomeric I-DmoI scaffold.

PubMed: 18974222
DOI: 10.1073/PNAS.0804795105

Experimental method

X-RAY DIFFRACTION (2.1 Å)