Rdh54 inhibits Rad54 mediated D-loop turnover.

Abstract

(A). Cartoon diagram illustrating the experimental design for monitoring Rad54 mediated D-loop maturation. The recipient DNA in this experiment is 21 nt of homologous DNA with 36 bp of non-homologous duplexed DNA (B). Representative agarose gels illustrating D-loop turnover for Rad51 alone, Rad51+Rad54 (30 nM), Rad51+Rad54+Rdh54 (10 nM), Rad51+Rad54+Rdh54 (30 nM), Rad51+Rad54+Rdh54 (90 nM), Rad51+Rdh54 (30 nM). (C). Quantification of D-loop turnover experiment as a Fraction of D-loop. The error bars represent the standard deviation of three independent experiments. (D). Normalized quantification of D-loop turnover. Experiments were normalized by setting the maximum fraction D-loop (10 min) to one and normalizing the other time points to that value. The error bars represent the standard deviation of three independent experiments. (E). Graphical depiction of fits of the D-loop turnover phase (10–45 min) of the reaction. The rates are represented from the slope of the best fit line for 0, 3:1, 1:1, and 1:3 molar ratios of Rad54:Rdh54.</p