摘要: DNA synthesis by polymerases (dPols) is central to duplication and maintenance of the genome in all living organisms. dPols catalyze formation a phosphodiester bond between incoming deoxynucleoside triphosphate terminal primer nucleotide with release pyrophosphate (PPi) group. It believed that an endergonic reaction PPi has be hydrolyzed accompanying pyrophosphatase enzymes ensure free energy change negative it can proceed forward direction. The fact proceeds vitro absence pyrophosphatases represents long-standing conundrum regarding thermodynamics reaction. Using time-resolved crystallography, we show hydrolysis intrinsic critical step catalyzed dPols. occurs after ensures energetically favorable without need for additional enzymes. Also, observe two Mg2+ ion assisted stepwise associative SN2 Overall, this study provides deep temporal insight primary enzymatic responsible duplication.
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