Rdh54 kinetically effects D-loop maturation on longer DNA templates.

Abstract

(A). Schematic diagram illustrating the longer fluorescently recipient DNA used in this experiments. The recipient DNA used in this experiment was 65 nt of homologous ssDNA and 25 bp of non-homologous dsDNA (B). Representative agarose gel representing a D-loop extension assay with Rad54 (30 nM) at 2, 5, 10, 20, 30, and 45 minutes. (C). Representative agarose gel representing D-loop extension assay with Rad54 (30 nM) + Rdh54 (30 nM) at 2, 5, 10, 20, 30, and 45 minutes. (D). Representative agarose gel representing D-loop extension assay with Rad54 (30 nM) + Rdh54K318R (30 nM) at 2, 5, 10, 20, 30, and 45 minutes. (E). Bar graph representing quantification of the fraction of D-loops that are extended at 10, 20, 30, and 45 min. The error bars represent the mean and standard deviation of 4–10 independent experiments. (F). A plot illustrating the reaction half-time generated from a single-phase association fit of the percentage D-loop extension. The crossbars represent the mean, and the error bars represent the upper and lower limit of the half-time based on the fit of the data.</p