Context dependence between subdomains in the DNA binding interface of the I-CreI homing endonuclease.

ABSTRACT: Homing endonucleases (HE) have emerged as precise tools for achieving gene targeting events. Redesigned HEs with tailored specificities can be used to cleave new sequences, thereby considerably expanding the number of targetable genes and loci. With HEs, as well as with other protein scaffolds, context dependence of DNA/protein interaction patterns remains one of the major limitations for rational engineering of new DNA binders. Previous studies have shown strong crosstalk between different residues and regions of the DNA binding interface. To investigate this phenomenon, we systematically combined mutations from three groups of amino acids in the DNA binding regions of the I-CreI HE. Our results confirm that important crosstalk occurs throughout this interface in I-CreI. Detailed analysis

SUBMITTER: Grizot S 

PROVIDER: S-EPMC3152339 | biostudies-literature | 2011 Aug

REPOSITORIES: biostudies-literature