DNA replication: In vitro single-molecule manipulation data analysis and models

DNA replication is a key biochemical process of the cell cycle. In the last years, analysis of in vitro single-molecule DNA replication events has provided new information that cannot be obtained with ensembles studies. Here, we introduce crucial techniques for the proper analysis and modelling of DNA replication in vitro single-molecule manipulation data. Specifically, we review some of the main methods to analyze and model the real-time kinetics of the two main molecular motors of the replisome: DNA polymerase and DNA helicase. Our goal is to facilitate access to and understanding of these techniques to promotetheir use in the study of DNA replication at the single-molecule level. A proper analysis of single-molecule data is crucial to obtain a detailed picture of, among others, the kinetics rates, equilibrium contants and conformational changes of the system under study. The techniques presented here have been used or can be adapted to study the operation of other proteins involved in nucleic acids metabolism. (C) 2021 The Authors. Published by Elsevier B.V. on behalf of Research Network of Computational and Structural Biotechnology.

Automated summary

The analysis and modeling techniques of in vitro single-molecule DNA replication events play a crucial role in understanding the process, with a focus on the real-time kinetics of DNA polymerase and DNA helicase. Proper analysis of single-molecule data is essential for obtaining a detailed picture of the system's dynamics, including kinetics rates, equilibrium constants, and conformational changes. These techniques can also be adapted to study the operation of other proteins involved in nucleic acids metabolism.