The link between restriction endonuclease fidelity and oligomeric state: A study with Bse634I |
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Authors: | Mindaugas Zaremba Giedrius Sasnauskas Virginijus Siksnys |
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Affiliation: | Institute of Biotechnology, Vilnius University, Graiciuno 8, LT-02241 Vilnius, Lithuania |
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Abstract: | Type II restriction endonucleases (REases) exist in multiple oligomeric forms. The tetrameric REases have two DNA binding interfaces and must synapse two recognition sites to achieve cleavage. It was hypothesised that binding of two recognition sites by tetrameric enzymes contributes to their fidelity. Here, we experimentally determined the fidelity for Bse634I REase in different oligomeric states. Surprisingly, we find that tetramerisation does not increase REase fidelity in comparison to the dimeric variant. Instead, an inherent ability to act concertedly at two sites provides tetrameric REase with a safety-catch to prevent host DNA cleavage if a single unmodified site becomes available. |
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Keywords: | REase, restriction endonuclease wt, wild-type SC, supercoiled OC, open-circular FLL, full-length linear FI, fidelity index BSA, bovine serum albumin DTT, dithiothreitol Tris, tris(hydroxymethyl)aminomethane EDTA, ethylenediaminetetraacetic acid |
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