Mutation of the Zip3 consensus phosphorylation sites by Mec1/Tel1 kinases alters its association with DSB sites and decreases crossover levels.

<p>(A) Meiotic progression of wild-type (ORD9670) or <i>zip3-4AQ</i> mutant (VBD1094) cells. Nuclear divisions were monitored by DAPI staining. (B) Zip3 association in the time-courses shown in (A) was monitored by ChIP with anti-Flag antibodies and revealed by qPCR with primer pairs covering the indicated regions. (C) Genetic distance in the <i>EST3-FAA3</i> interval on chromosome IX measured in the <i>ZIP3-</i>Flag (VBD1229) and <i>zip3-4AQ-</i>Flag (VDB1113) strains (see also <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003416#pgen.1003416.s016" target="_blank">Table S2</a>). The configuration of the hemizygous resistance markers used to measure the genetic distances is shown in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003416#pgen.1003416.s008" target="_blank">Figure S8</a>. (D) Genetic distances determined for nine intervals distributed on three chromosomes. See also <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003416#pgen.1003416.s015" target="_blank">Table S1</a>. <i>ZIP3</i>-Flag: VBH334/VBH335 strain; <i>zip3-4AQ</i>-Flag: VBH332/VBH331 strain. (E) Physical analysis of COs in the <i>EST3-FAA3</i> interval in <i>ZIP3</i>, <i>zip3-4AQ</i> and <i>mus81Δ</i> mutants. Genomic DNA was extracted at the indicated times of synchronous meiosis and digested with <i>Bsp</i>EI and <i>Bss</i>HII. The parental (P1 and P2) and CO (R1 and R2) bands are indicated. The R2 band was quantitated and expressed as % of total DNA. <i>ZIP3</i>: VBD1229; <i>zip3-4AQ</i>: VBD1113; <i>ZIP3 mus81Δ</i>: VBD1244; <i>zip3-4AQ mus81Δ</i>: VBD1245. The graph indicates the mean of two independent experiments. Error bars represent standard deviation.