RecA and DNA recombination: a review of molecular mechanisms

Elsa del Val (1, 2) , William Nasser (1, 2) , Hafid Abaibou , Sylvie Reverchon (1, 2)

1 CRP - Chromatine et Régulation de la Pathogénie bactérienne
2 MAP - Microbiologie, adaptation et pathogénie

Recombinases are responsible for homologous recombination and maintenance of genome integrity. In Escherichia coli, the recombinase RecA forms a nucleoprotein filament with the ssDNA present at a DNA break and searches for a homologous dsDNA to use as a template for break repair. During the first step of this process, the ssDNA is bound to RecA and stretched into a Watson-Crick base-paired triplet conformation. The RecA nucleoprotein filament also contains ATP and Mg 2+ , two cofactors required for RecA activity. Then, the complex starts a homology search by interacting with and stretching dsDNA. Thanks to supercoiling, intersegment sampling and RecA clustering, a genome-wide homology search takes place at a relevant metabolic timescale. When a region of homology 8 to 20 base pairs in length is found and stabilized, DNA strand exchange proceeds, forming a heteroduplex complex that is resolved through a combination of DNA synthesis, ligation and resolution. RecA activities can take place without ATP hydrolysis, but this latter activity is necessary to improve and accelerate the process. Protein flexibility and monomer-monomer interactions are fundamental for RecA activity, which functions cooperatively. A structure/function relationship analysis suggests that the recombinogenic activity can be improved and that recombinases have an inherently large recombination potential. Understanding this relationship is essential for designing RecA derivatives with enhanced activity for biotechnology applications. For example, this protein is a major actor in the recombinase polymerase isothermal amplification (RPA) used in point-of-care diagnostics.

Domaines: Biologie moléculaire

Elsa del Val, William Nasser, Hafid Abaibou, Sylvie Reverchon. RecA and DNA recombination: a review of molecular mechanisms. Biochemical Society Transactions, 2019, ⟨10.1042/BST20190558⟩. ⟨hal-02320683⟩