FIGURE 5.

Rad59 alleviates the inhibitory effect of Rad51 in Rad52-mediated DNA annealing. Experiments were carried out as described under “Experimental Procedures.” RPA-complexed ssDNA was preincubated without or with Rad51 (134 nm), and the reactions were initiated by the addition of Rad52 (40 nm) and/or Rad59 (40 or 80 nm). The reactions either lacking or containing Rad51 are shown in black and blue, respectively. The reaction containing Rad52 alone (black open squares), Rad51 followed by Rad52 (blue open triangles), Rad51 followed by Rad52 and 40 nm Rad59 (blue solid triangles), and Rad51 followed by Rad52 and 80 nm Rad59 (blue diamonds) are shown. The results are the averages obtained from at least three independent experiments, and the error bars represent one standard deviation. Control reactions containing Rad59 only (80 nm; green solid circles), Rad51 and Rad59 (green open circles), and Rad52 and Rad59 (black solid squares) are shown. The results are the averages obtained from at least two independent experiments, and the error bars represent the variation (where absent, the error bars are smaller than the symbol).