2I3P: K28r Mutant Of Homing Endonuclease I-crei

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.
PDB ID: 2I3PDownload
MMDB ID: 53473
PDB Deposition Date: 2006/8/20
Updated in MMDB: 2017/10
Experimental Method:
x-ray diffraction
Resolution: 2.3  Å
Source Organism:
Chlamydomonas reinhardtii
Similar Structures:
Biological Unit for 2I3P: tetrameric; determined by author
Molecular Components in 2I3P
Label Count Molecule
Proteins (2 molecules)
DNA Endonuclease I-crei
Molecule annotation
Nucleotides(2 molecules)
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'
Molecule annotation
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'
Molecule annotation
Chemicals (2 molecules)
* Click molecule labels to explore molecular sequence information.

Citing MMDB