Rad51 is the central catalyst of homologous recombination in eukaryotes and is thus critical for maintaining genomic integrity. Recent crystal structures of filaments formed by Rad51 and the closely related archeal RadA and eubacterial RecA proteins place the ATPase site at the protomeric interface. To test the relevance of this feature, we mutated conserved residues at this interface and examined their effects on key activities of Rad51: ssDNA-stimulated ATP hydrolysis, DNA binding, polymerization on DNA substrates and catalysis of strand-exchange reactions. Our results show that the interface seen in the crystal structures is very important for nucleoprotein filament formation. H352 and R357 of yeast Rad51 are essential for assembling the catalytically competent form of the enzyme on DNA substrates and coordinating its activities. However, contrary to some previous suggestions, neither of these residues is critical for ATP hydrolysis.

Adenosine Triphosphatases, Adenosine Triphosphate, Amino Acid Substitution, Atomic Force, DNA, DNA binding, DNA repair, Microscopy, Mutagenesis, Nucleotides, Protein Subunits, Rad51 Recombinase, Rad51 protein, Saccharomyces cerevisiae Proteins, Single-Stranded, Site-Directed, adenosine triphosphatase, article, catalysis, controlled study, crystal structure, double stranded DNA break, enzyme activity, mutational analysis, nonhuman, priority journal, protein analysis, protein expression, protein hydrolysis, protein interaction, protein polymerization, structure activity relation, yeast
dx.doi.org/10.1093/nar/gkn973, hdl.handle.net/1765/15747
Nucleic Acids Research
Erasmus MC: University Medical Center Rotterdam

Grigorescu, A.A, Vissers, J.H.A, Ristic, D, Pigli, Y.Z, Lynch, T.W, Wyman, C, & Rice, P.A. (2009). Inter-subunit interactions that coordinate Rad51's activities. Nucleic Acids Research, 37(2), 557–567. doi:10.1093/nar/gkn973