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

Rebeca Bocanegra; G A Ismael Plaza; Carlos R Pulido; Borja Ibarra

doi:10.1016/j.csbj.2021.04.013

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

Comput Struct Biotechnol J. 2021 Apr 20:19:2057-2069. doi: 10.1016/j.csbj.2021.04.013. eCollection 2021.

Authors

Rebeca Bocanegra¹, G A Ismael Plaza¹, Carlos R Pulido¹, Borja Ibarra¹

Affiliation

¹ IMDEA Nanociencia, Faraday 9, Campus Cantoblanco, 28049 Madrid, Spain.

Abstract

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.

Keywords: DNA replication; Fluorescence spectroscopy; Force spectroscopy; Replisome; Single-molecule.

Publication types

Review