Homologous Recombination and DNA Intermediates Analyzed by Electron Microscopy

Clara Basto, Eliana Moreira-Tavares, Ali Akbar Muhammad, Sonia Baconnais, Gerard Mazón, Eric Le Cam, Pauline Dupaigne

Résultats de recherche: Contribution à un journalArticleRevue par des pairs

Résumé

Homologous recombination (HR) is a high-fidelity DNA repair pathway that uses a homologous DNA sequence as a template. Recombinase proteins are the central HR players in the three kingdoms of life. RecA/RadA/Rad51 assemble on ssDNA, generated after the processing of double-strand breaks or stalled replication forks into an active and dynamic presynaptic helical nucleofilament. Presynaptic filament formation is regulated by a series of partners of the recombinase, such as scRad52/hBRCA2 mediators or anti-recombinase proteins, to form an active machinery involved in homology search, pair-matching, and invasion within homologous sequences. During homology search, but also during strand invasion, the multiprotein complexes that form the nucleofilament induce the formation of a variety of DNA intermediate states. Here we present specific approaches to study and characterize the different DNA and DNA-protein intermediates formed during homologous recombination. The combination of powerful electron microscopy and sample preparation methods provides a better understanding of these proteins' molecular activity and their interactions.

langue originaleAnglais
Pages (de - à)239-257
Nombre de pages19
journalMethods in molecular biology (Clifton, N.J.)
Volume2881
Les DOIs
étatPublié - 1 janv. 2025
Modification externeOui

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