Molina…Montoya {Nat Struct Mol Biol 22: 65}
The enzyme I-DmoI (surface model in purple) is specifically associated to the double strand of DNA (stick model, yellow and green) before the catalysis of an specific doble strand break. One of the crystal X-ray diffraction pattern is used as background.
Visualizing phosphodiester-bond hydrolysis by an endonuclease
Molina R, Stella S, Redondo P, Gomez H, Marcaida MJ, Orozco M, Prieto J, Montoya G.
Nat Struct Mol Biol. 2015 Jan; 22: 65. Epub 2014 Dec 8.
PubMed: 25486305. Doi: 10.1038/nsmb.2932
TThe enzymatic hydrolysis of DNA phosphodiester bonds has been widely studied, but the chemical reaction has not yet been observed. Here we follow the generation of a DNA double-strand break (DSB) by the Desulfurococcus mobilis homing endonuclease I-DmoI, trapping sequential stages of a two-metal-ion cleavage mechanism. We captured intermediates of the different catalytic steps, and this allowed us to watch the reaction by ‘freezing’ multiple states. [….]
Additional Material
- Detailed video depicting the active center during double strand break catalysis.
- Video showing the phosphodiester catalysis and the possible applications for an engineered enzyme.
