An internal deletion mutant stabilized Rev1 protein in the S phase.

Abstract

<p><b>A,</b> Schematic diagram of the fission yeast Rev1 domain structure. Parallelogram, BRCT; rectangle, Y family-conserved region; ellipse, ubiquitin binding motif; hexagon, Lys-rich region. Rev1dK and Rev1dKK are internal deletion mutants lacking the amino acids from 761 to 818 and 761 to 797, respectively. <b>B,</b> The Rev1dKK mutant stabilized Rev1 protein. Flag-tagged <i>rev1^wt</i> and <i>rev1dKK</i> cells were grown, and whole cell extracts were prepared by the boiling method. The upper panel shows a western blot of Rev1 protein, and the lower panel shows CBB staining of the membrane as a loading control. The left lane represents Rev1<i>^wt</i>, and the right lane represents Rev1dKK. <b>C,</b> Rev1dKK protein was stable in S phase. Time course samples of the <i>rev1dKK^flag cdc25</i> strain were prepared. The samples were taken every 30 min after the release. The lower panels show the expression patterns of Rev1dKK, Cdc13, and Cdc2.</p