2026-06-28T20:39:10Zhttps://docta.ucm.es/rest/oai/request
oai:docta.ucm.es:20.500.14352/1054392025-09-18T16:54:20Zcom_20.500.14352_14col_20.500.14352_15
Docta Complutense author Morin, Jose A. author Cao García, Francisco Javier author Lazaro, Jose M. author Ricardo Arias-Gonzalez, J. author Valpuesta, Jose M. author Carrascosa, Jose L. author Salas, Margarita author Ibarra, Borja 2024-07-02T10:00:27Z 2024-07-02T10:00:27Z 2012 Morin, J. A., Cao, F. J., Lázaro, J. M., Arias-Gonzalez, J. R., Valpuesta, J. M., Carrascosa, J. L., ... & Ibarra, B. (2012). Active DNA unwinding dynamics during processive DNA replication. Proceedings of the National Academy of Sciences, 109(21), 8115-8120. 0027-8424 10.1073/pnas.1204759109 https://hdl.handle.net/20.500.14352/105439 1091-6490 https://doi.org/10.1073/pnas.1204759109 https://www.pnas.org/doi/full/10.1073/pnas.1204759109 Duplication of double-stranded DNA (dsDNA) requires a fine-tuned coordination between the DNA replication and unwinding reactions. Using optical tweezers, we probed the coupling dynamics between these two activities when they are simultaneously carried out by individual Phi29 DNA polymerase molecules replicating a dsDNA hairpin. We used the wild-type and an unwinding deficient polymerase variant and found that mechanical tension applied on the DNA and the DNA sequence modulate in different ways the replication, unwinding rates, and pause kinetics of each polymerase. However, incorporation of pause kinetics in a model to quantify the unwinding reaction reveals that both polymerases destabilize the fork with the same active mechanism and offers insights into the topological strategies that could be used by the Phi29 DNA polymerase and other DNA replication systems to couple unwinding and replication reactions. eng Active DNA unwinding dynamics during processive DNA replication journal article 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