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. 2008 Jul 14;182(1):27–33. doi: 10.1083/jcb.200712080

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© 2008 Tan et al.

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 5. — An essential role of mars is to promote dTACC dephosphorylation. (A) Mars coprecipitates dTACC and Msps from D. melanogaster embryonic nuclear extracts. Immunoprecipitation of arm-GAL4 (−) and arm>mars^WT (+) embryo extracts with Myc antibody, followed by immunoblotting with dTACC and Msps antibodies, showed binding to FM-tagged Mars. Immunoprecipitation with dTACC antibody followed by immunoblotting with Mars antibody confirmed dTACC binding. (B) Distribution of dTACC and p-dTACC on mitotic spindles in embryos from mothers of different genotypes, as indicated. In wild type, p-dTACC is only found at the centrosome but in mars mutants, p-dTACC abnormally accumulates on mitotic spindles. Total dTACC staining is largely unaffected in mars mutants. A nonphosphorylatable mutant form of dTACC (dTACC^SL), but not wild-type dTACC (dTACC^WT), restores spindle structure and normal distribution of p-dTACC in a mars^1/P mutant background. Similarly, arm>mars^WT, but not arm>mars^FA, restores normal spindle structure and p-dTACC staining in a mars¹ background. Bar, 10 μm. (C) Linescans of fluorescence intensity (arbitrary units) across spindles from embryos of different genotypes, as indicated. The distribution of p-dTACC (red trace) is shown relative to α-tubulin (green trace). (D) Top graph shows quantification of ratio of spindle/centrosomal p-dTACC staining. dTACC^SL, but not dTACC^WT, restores a normal p-dTACC ratio in embryos from mars^1/P mothers. Bottom graph shows that dTACC^SL, but not dTACC^WT, rescues lethality of embryos laid by mars^1/P mothers. Hatch ratios are plotted as the mean ± standard error from n = 5 experiments.