Redesign of the monomer-monomer interface of Cre recombinase yields an obligate heterotetrameric complex.

ABSTRACT: Cre recombinase catalyzes the cleavage and religation of DNA at loxP sites. The enzyme is a homotetramer in its functional state, and the symmetry of the protein complex enforces a pseudo-palindromic symmetry upon the loxP sequence. The Cre-lox system is a powerful tool for many researchers. However, broader application of the system is limited by the fixed sequence preferences of Cre, which are determined by both the direct DNA contacts and the homotetrameric arrangement of the Cre monomers. As a first step toward achieving recombination at arbitrary asymmetric target sites, we have broken the symmetry of the Cre tetramer assembly. Using a combination of computational and rational protein design, we have engineered an alternative interface between Cre monomers that is functional yet incom

SUBMITTER: Zhang C 

PROVIDER: S-EPMC4605323 | biostudies-literature | 2015 Oct

REPOSITORIES: biostudies-literature