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 singlemolecule 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 DN...

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Detalles Bibliográficos
Autores: Jarillo, Javier, Ibarra, Borja, Cao García, Francisco Javier
Tipo de recurso: artículo
Fecha de publicación:2021
País:España
Institución:Universidad Complutense de Madrid (UCM)
Repositorio:Docta Complutense
Idioma:inglés
OAI Identifier:oai:docta.ucm.es:20.500.14352/104481
Acceso en línea:https://hdl.handle.net/20.500.14352/104481
Access Level:acceso abierto
Palabra clave:57.088.7
DNA replication
DNA unwinding
DNA polymerase
Helicase
Single-molecule
Real-time kinetics
Biotecnología
2409.02 Ingeniería Genética
Descripción
Sumario:DNA replication is a key biochemical process of the cell cycle. In the last years, analysis of in vitro singlemolecule 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.