In crystallo observation of active site dynamics and transient metal ion binding within DNA polymerases

DNA polymerases are the enzymatic catalysts that synthesize DNA during DNA replication and repair. Kinetic studies and x-ray crystallography have uncovered the overall kinetic pathway and led to a two-metal-ion dependent catalytic mechanism. Diffusion-based time-resolved crystallography has permitted the visualization of the catalytic reaction at atomic resolution and made it possible to capture transient events and metal ion binding that have eluded static polymerase structures. This review discusses past static structures and recent time-resolved structures that emphasize the crucial importance of primer alignment and different metal ions binding during catalysis and substrate discrimination.

Automated summary

DNA polymerases are essential enzymes for DNA replication and repair, and they operate through a two-metal-ion dependent catalytic mechanism. Recent advances in diffusion-based time-resolved crystallography have allowed the atomic-resolution visualization of catalytic reactions, revealing the importance of primer alignment and different metal ions binding during catalysis and substrate discrimination. This review highlights the significance of both static and time-resolved structures in understanding DNA polymerase function.