Eso1 and Rev1 were associated in G₁ phase.

Abstract

<p><b>A,</b><i>eso1^Δpolh</i> and <i>rev1Δ</i> mutants exhibited similar sensitivities to UV irradiation. Cells in the logarithmic growth phase were serially diluted by 5 fold. Cells were then spotted on YES plates and exposed to UV light (0, 100, or 150 J/m<i>²</i>). Plates were incubated at 30°C, and the growth of wt, <i>eso1^Δpolh</i>, and <i>rev1Δ</i> strains was observed 3 days after the irradiation. <b>B,</b> Eso1 associated with Rev1 in <i>cdc10</i>-arrested extracts. <i>cdc10 rev1^flag eso1^V5</i> and <i>cdc25 rev1^flag eso1^V5</i> strains were first grown at 25°C, and the cultures were split into two. The first half was further grown at 25°C, and the second half was grown at 36°C for 3 h. The cells were harvested, and whole cell extracts were prepared. Immunoprecipitation was then carried out using anti-flag or anti-V5 antibodies. The panels represent input, Flag IP, and V5 IP of Rev1 and Eso1 protein in the <i>cdc10</i> strain at 25°C, the <i>cdc10</i> strain at 36.5°C, the <i>cdc25</i> strain at 25°C, and the <i>cdc25</i> strain at 36.5°C. <b>C,</b> Eso1 was coprecipitated with Rev7. <i>eso1^myc</i> and <i>eso1^myc rev7^flag</i> strains were grown, and whole cell extracts were prepared. Immunoprecipitation was performed using anti-flag antibodies. The panels show the expression of Eso1 and Rev7 in input samples and immunoprecipitated fractions. <b>D,</b> Rev7 failed to coprecipitate Eso1 in the absence of Rev1. <i>eso1^myc rev7^flag</i> and <i>eso1^myc rev7^flag rev1Δ</i> strains were grown, and whole cell extracts were prepared. Immunoprecipitation was performed using anti-flag antibodies. The panels show the expression of Eso1 and Rev7 in input samples and immunoprecipitated fractions.</p