|Cooperative dynamics of a DNA polymerase replicating complex|Moors, C.; Herdewijn, P.; Robben, J.; Ceulemans, A. (2013). Cooperative dynamics of a DNA polymerase replicating complex. Biochimica et Biophysica Acta-Proteins and Proteomics 1834(12): 2554-2563. dx.doi.org/10.1016/j.bbapap.2013.09.003
In: Biochimica et Biophysica Acta-Proteins and Proteomics. ELSEVIER SCIENCE BV: Amsterdam. ISSN 1570-9639, more
Replica exchange with flexible tempering; DNA polymerase; Protein-DNAinteraction; Therrnococcus gorgonarius
|Authors|| || Top |
- Moors, C.
- Herdewijn, P.
- Robben, J.
- Ceulemans, A.
Engineered DNA polymerases continue to be the workhorses of many applications in biotechnology, medicine and nanotechnology. However, the dynamic interplay between the enzyme and the DNA remains unclear. In this study, we performed an extensive replica exchange with flexible tempering (REFT) molecular dynamics simulation of the ternary replicating complex of the archaeal family B DNA polymerase from the thermophile Thermococcus gorgonarius, right before the chemical step. The convoluted dynamics of the enzyme are reducible to rigid-body motions of six subdomains. Upon binding to the enzyme, the DNA double helix conformation changes from a twisted state to a partially untwisted state. The twisted state displays strong bending motion, whereby the DNA oscillates between a straight and a bent conformation. The dynamics of double-stranded DNA are strongly correlated with rotations of the thumb toward the palm, which suggests an assisting role of the enzyme during DNA translocation. In the complex, the primer-template duplex displays increased preference for the B-DNA conformation at the n - 2 and n - 3 dinucleotide steps. Interactions at the primer 3 ' end indicate that Thr541 and Asp540 are the acceptors of the first proton transfer in the chemical step, whereas in the translocation step both residues hold the primer 3 ' terminus in the vicinity of the priming site, which is crucial for high processivity.