Single strand DNA binding and annealing activities in the yeast recombination factor Rad59

Saccharomyces cerevisiae RAD59 gene is required for homologous recombination processes and normal level of resistance to ionizing radiation. To study the biochemical functions of Rad59, it was overproduced in yeast and purified to near homogeneity. Rad59 binds DNA, showing much higher affinity for ssDNA than dsDNA. Rad59 also anneals complementary DNA strands, and order of addition experiments indicate that maximal annealing efficiency is achieved when both complementary DNA strands are present upon addition of Rad59. Thus, Rad59 resembles its homolog Rad52 in being able to bind ssDNA and anneal complementary DNA strands. However, unlike Rad52, DNA annealing by Rad59 is not accelerated by the ssDNA binding factor RPA. DNA binding and strand annealing are likely to be important for the biological functions of Rad59 in general recombination and in the single-strand annealing pathway of recombination.