FokI requires two specific DNA sites for cleavage.
作者： Éva Scheuring Vanamee , Sandro Santagata , Aneel K Aggarwal
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摘要: FokI is a bipartite restriction endonuclease that recognizes non-palindromic DNA sequence, and then makes double-stranded cuts outside of sequence to leave 5' overhang. Earlier kinetic crystallographic studies suggested might function as dimer. Here, we show, using dynamic light-scattering, gel-filtration analytical ultracentrifugation, dimerizes only in the presence divalent metal ions. Furthermore, analysis DNA-bound complex reveals two copies recognition are incorporated into dimeric formation this essential for full activation cleavage. These results have broad implications mechanism by which monomeric type II endonucleases achieve high fidelity.
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