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. 2008 Sep 8;182(5):873–883. doi: 10.1083/jcb.200801054

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© 2008 Queralt and Uhlmann

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 4. — Zds1 and the MEN. (A) The essential MEN kinase Cdc15 is not required for Zds1-induced Cdc14 release. Strains Y3288 (MATa MET-CDC20 GAL-Flag-ZDS1 CDC14-Pk₉) and Y1429 (as Y3288, but cdc15-2) were arrested in metaphase by Cdc20 depletion and shifted to 37°C for 60 min before Zds1 induction. (B) cdc15-2 and zds1Δ zds2Δ mutations display a synthetic growth defect. Strains Y699 (W303), Y3196 (MATa cdc15-2), Y3117 (MATa zds1Δ zds2Δ), and Y3428 (as Y3117, but cdc15-2) were streaked onto YPD plates and incubated at 31°C for 2 d. (C) Zds1 overexpression rescues growth of cdc15-2 cells at restrictive temperature. Strains Y3196 and Y3191 (MATa cdc15-2 GAL1-ZDS1) were streaked onto YP plates containing raffinose and galactose, to induce Zds1 expression, at 33°C for 2 d. (D) Cdc42 effectors and PAK-kinases are not required for Zds1-induced Cdc14 release. Strains Y3288 (MATa MET-CDC20 GAL-Flag-ZDS1 CDC14-Pk₉), Y1409 (as Y3288, but ste20Δ), Y1410 (as Y3288, but cla4Δ), Y1412 (as Y3288, but gic1Δ), Y1411 (as Y3288, but gic2Δ), and Y1414 (as Y3288, but gic1Δ gic2Δ) were arrested in metaphase by Cdc20 depletion and Zds1 expression was induced. Cdc14 nucleolar release was monitored by immunofluorescence. Equal Zds1 expression levels were confirmed by Western blotting (not depicted).