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DNA replication machinery: Insights from in vitro single-molecule approaches

Journal

COMPUTATIONAL AND STRUCTURAL BIOTECHNOLOGY JOURNAL
Volume 19, Issue -, Pages 2057-2069

Publisher

ELSEVIER
DOI: 10.1016/j.csbj.2021.04.013

Keywords

DNA replication; Single-molecule; Force spectroscopy; Fluorescence spectroscopy; Replisome

Funding

  1. Spanish Ministry of Economy and Competitiveness [PGC2018099341BI00, PRE2019-088885]
  2. Comunidad de Madrid [PEJD2016_IND2451, PEJD2018PRE_SAL8038, P2018INMT4321]
  3. 'Severo Ochoa' Program for Centers of Excellence in RD (MINECO) [SEV20160686]

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The study reveals that the replisome machinery is stochastic, versatile, and highly dynamic, with transient protein-protein and protein-DNA interactions playing a key role in robust DNA replication.
The replisome is the multiprotein molecular machinery that replicates DNA. The replisome components work in precise coordination to unwind the double helix of the DNA and replicate the two strands simultaneously. The study of DNA replication using in vitro single-molecule approaches provides a novel quantitative understanding of the dynamics and mechanical principles that govern the operation of the replisome and its components. 'Classical' ensemble-averaging methods cannot obtain this information. Here we describe the main findings obtained with in vitro single-molecule methods on the performance of individual replisome components and reconstituted prokaryotic and eukaryotic replisomes. The emerging picture from these studies is that of stochastic, versatile and highly dynamic replisome machinery in which transient protein-protein and protein-DNA associations are responsible for robust DNA replication. (C) 2021 The Author(s). Published by Elsevier B.V. on behalf of Research Network of Computational and Structural Biotechnology.

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