Rad54-2A is not functional <i>in vivo</i> and is expressed at normal protein levels.

<p><b>A.</b>. <i>rad54-2A</i> cells are extremely MMS-sensitive. Cell cultures of wild type (WDHY2217), <i>rad54-Y494A, F495A ( = rad54-2A</i>, WDHY2625), and <i>rad54-Δ</i> (WDHY2571) were grown in YPD, diluted to OD₆₀₀ of 1, followed by 6 five-fold serial dilutions. Cells were plated onto YPD containing 0.0025% or 0.01% MMS and grown at 30°C for 2 days and images were acquired to document cell growth. <b>B.</b> Comparison of endogenous wild type Rad54 and Rad54-2A protein levels. Extracts (100 µg) from wild type (WDHY2217), <i>rad54-2A</i> (WDHY2625) and <i>rad54-Δ</i> (WDHY2571) cells were immunoblotted with anti-<i>S. cerevisiae</i> Rad54 serum. The Rad54 protein levels were quantified using a crossreacting band as loading control. The ratios of the intensity of Rad54/control are indicated.