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Updated: Nov 9, 2025
Author Spotlight: Investigating the Motion Dynamics of the Eukaryotic Replisome Components at the Single-Molecule Level
Published on: July 26, 2024
Single-Molecule Tethered Particle Motion Studies on the DNA Recombinase Filament Assembly and Disassembly
Chih-Hao Lu1,2, Wei-Hsuan Lan1, Hung-Wen Li3
1Department of Chemistry, National Taiwan University, Taipei, Taiwan.
Abstract:
Bacterial RecA and eukaryotic Rad51 are recombinases indispensable for DNA homologous recombination and repair of double-stranded DNA breaks. Understanding the functions and biophysical properties of the DNA recombinases benefits the research in human medicine such as cancer biology. Single-molecule techniques provide the mechanistic details of complex biological reactions. Tethered particle motion (TPM) experiment is a simple and multiplex single-molecule tool to monitor DNA-protein interactions. We have developed a single-molecule TPM assay to study DNA recombinase filament assembly and disassembly on individual DNA molecules in real time. Characterization of the temporal change of the Brownian motion of DNA tethers during recombinase assembly and disassembly in real time allows the determination of multiple kinetic parameters of nucleation rate, extension rate, dissociation rate, and length of the recombinase-DNA filament.
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