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. 2011 Jun 27;193(7):1213–1228. doi: 10.1083/jcb.201103076

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© 2011 Bizzari and Marston

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.rupress.org/terms). 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 1. — Impaired nuclear division, random chromosome segregation, and ectopic Cdc14 release in cdc55mn cells. (A–D) Meiosis was induced in strains carrying heterozygous CEN5-GFP, PDS1-18MYC, and otherwise wild type (AM4796), cdc55mn (AM4891), or cdc55Δ (AM5338). The percentages of binucleate and tetranucleate cells (n = 200; A–C) or the pattern of GFP foci in binucleate cells (n > 800; D) is shown for a representative experiment. (E) Wild-type (AM6040) and cdc55mn (AM5936) strains carrying homozygous CEN5-GFP were analyzed as described in D. (F and G) Wild-type (AM6633) and cdc55mn (AM6626) cells carrying 3HA-CDC14, GAL-NDT80, and pGPD1-GAL4(848).ER were induced to sporulate and released from a pachytene block at 6 h. The percentages of cells with the indicated spindle morphology and with Cdc14 released from the nucleolus are shown for a representative experiment. (H–K) Wild-type (AM6935) and cdc55mn (AM6942) cells carrying CDC14-GFP, GFP-TUB1, and homozygous tetR-tdTomato were filmed. (H–J) Still images from Videos 1 (wild-type; H), 2 (cdc55mn; I), and 3 (cdc55mn; J). Arrows indicate Cdc14 sequestered in the nucleolus. Bars, 1 µm. (K) Behavior of cdc55mn cells that were in prophase I (as judged by spindle morphology) at the start of filming and released Cdc14 from the nucleolus (563/1,203 prophase I cells). Examples of extruding microtubules are shown in fixed cells in Fig. S2. Times are given in minutes. (L–N) The time elapsed between the first Cdc14 release to Cdc14 resequestration (L), the second Cdc14 release to nuclear division (M), or Cdc14 resequestration and rerelease (N) are shown for cells that were in prophase I at the start of filming and in which two rounds of Cdc14 release were observed (wild type, n = 95; cdc55mn, n = 80). Box boundaries represent the bottom quartile and top quartile. The red line indicates the median, diamonds indicate the mean, and error bars represent the minimum and maximum values observed except for in the case of N in which two outliers (triangles) were excluded from the analysis. Because images were captured at 10-min intervals, the time elapsed between two observed events is subject to an error of 20 min.