Rev1 protein was stabilized under the restrictive condition for the <i>mts2</i> or <i>mts3</i> temperature-sensitive mutant.

Abstract

<p><b>A,</b> Protein level of Rev1 was increased after the temperature shift in the <i>mts2</i> temperature-sensitive mutant. The <i>mts2-U31</i> mutant, harboring-flag tagged Rev1, was first grown at 25°C, and the temperature was then shifted to 36.5°C. Samples were collected every 2 h until 4 h after the shift. Whole cell extracts were prepared. The panels show the protein expression of Rev1, Cdc13, Cdc2, and Pcn1. <b>B,</b> The protein level of Rev1dK did not increase at the restrictive temperature of <i>mts2-U31</i>. The panels show the protein expression of Rev1dK, Cdc13, and Cdc2 in the <i>mts2-U31</i> strain, and that of Rev1dK and Cdc2 in the <i>mts2^wt</i> control. The lanes represent the protein expression at 0, 2, and 4 h after the temperature shift in the <i>mts2-U31 or mts2^wt</i> strain. <b>C,</b> The protein level of Rev1 increased after the temperature shift in the <i>mts3</i> temperature-sensitive mutant. The <i>mts3-U32</i> mutant, harboring flag-tagged Rev1, was first grown at 25°C, and the temperature was then shifted to 36.5°C. The samples were collected every 2 h until 6 h after the shift. Whole cell extracts were prepared. The panels show the protein expression of Rev1 and Cdc2, as well as CBB staining of the membrane. <b>D,</b> The protein level of Rev1dK did not increase at the restrictive temperature in the <i>mts3-U32</i> strain. The panels show the protein expression of Rev1dK and Cdc2, as well as CBB staining of the membrane. The lanes represent the protein expression at 0, 2, 4, and 6 h after the temperature shift in the <i>mts3-U32</i> strain.</p