DNA polymerase conformational dynamics and the role of fidelity-conferring residues: Insights from computational simulations

Frontiers in Molecular Biosciences

卷:3

Giorgio eColombo¹  Massimiliano eMeli¹  Marko eSustarsic²  Timothy D Craggs²  Achillefs N Kapanidis² 

[1] National Research Council of Italy;

[2] University of Oxford;

关键词: DNA Polymerase I;    allostery;    molecular recognition;    molecular dynamics simulations;    replication fidelity;   

DOI  :  10.3389/fmolb.2016.00020

来源: DOAJ

【 摘 要 】

Herein we investigate the molecular bases of DNA polymerase I conformational dynamics that underlie the replication fidelity of the enzyme. Such fidelity is determined by conformational changes that promote the rejection of incorrect nucleotides before the chemical ligation step. We report a comprehensive atomic resolution study of wild type and mutant enzymes in different bound states and starting from different crystal structures, using extensive molecular dynamics (MD) simulations that cover a total timespan of ~ 5 microseconds. The resulting trajectories are examined via a combination of novel methods of internal dynamics and energetics analysis, aimed to reveal the principal molecular determinants for the (de)stabilization of a certain conformational state. Our results show that the presence of fidelity-decreasing mutations or the binding of incorrect nucleotides in ternary complexes tend to favor transitions from closed towards open structures, passing through an ensemble of semi-closed intermediates. The latter ensemble includes the experimentally observed ajar conformation which, consistent with previous experimental observations, emerges as a molecular checkpoint for the selection of the correct nucleotide to incorporate. We discuss the implications of our results for the understanding of the relationships between the structure, dynamics and function of DNA polymerase I at the atomistic level.

【 授权许可】

Unknown