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Fig. 1. Molecular lesions, TAC-1 and ZYG-9 distribution in five new zyg-9 temperature-sensitive mutant alleles. (A) Schematic representation of ZYG-9 (1-1415) showing the position and nature of the mutations identified in the five new zyg-9 alleles. Yellow boxes, TOG domains; line above the protein, TAC-1-binding region TBR (thick line, strong binding; thin line, weak binding) (Bellanger and Gönczy, 2003). (B-M) One-cell-stage embryos during mitosis in the wild-type (B,C) or zyg-9 (D-M) temperature-sensitive mutant alleles shifted at 25°C for >12 hours and stained with antibodies against ZYG-9 (B,D,F,H,J,L) or TAC-1 (C,E,G,I,K,M) and -tubulin. Left panels show ZYG-9 or TAC-1 staining alone, right panels the merge of ZYG-9 or TAC-1 (red), -tubulin (green) and DNA (blue). The histograms on the right of each staining series represent the quantification of ZYG-9 and TAC-1 levels in the cytoplasm (white bars) and the ratio between the levels of ZYG-9 or TAC-1 at centrosomes and in the cytoplasm (black bars) (Materials and Methods). Values are given as the mean ± s.e.m.; a.u., arbitrary units relative to the wild-type values, which have been set to 1 (dashed lines). Numbers of embryos analyzed for TAC-1 and ZYG-9 were: wild type, 17 and 13; zyg-9(or593), 10 and 9; zyg-9(or623), 10 and 13; zyg-9(or628), 10 and 8; zyg-9(or634), 11 and 10; zyg-9(or635), 10 and 10. Anterior is left, posterior is right; bar, 10 µm. Images in panels D, E and K are maximum-intensity projection of two 1-µm-thick confocal sections; the remaining images are single 1-µm-thick confocal sections. We found also that the distribution of ZYG-9 and TAC-1 is comparable with that of the wild type in zyg-9(or623), zyg-9(or634) and zyg-9(or635) embryos at 15°C (data not shown).

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