Structure of the I-SceI nuclease complexed with its dsDNA target and three catalytic metal ions.

ABSTRACT: Homing endonucleases are highly specific DNA-cleaving enzymes that recognize and cleave long stretches of DNA. The engineering of these enzymes provides instruments for genome modification in a wide range of fields, including gene targeting. The homing endonuclease I-SceI from the yeast Saccharomyces cerevisiae has been purified after overexpression in Escherichia coli and its crystal structure has been determined in complex with its target DNA. In order to evaluate the number of ions that are involved in the cleavage process, thus determining the catalytic mechanism, crystallization experiments were performed in the presence of Mn(2+), yielding crystals that were suitable for X-ray diffraction analysis. The crystals belonged to the orthorhombic space group P212121, with unit-cell paramete

SUBMITTER: Prieto J 

PROVIDER: S-EPMC4909248 | biostudies-literature | 2016 Jun

REPOSITORIES: biostudies-literature