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Fig. 1. Mcp6 is a meiosis-specific coiled-coil protein that harbours three homologous regions conserved in other proteins that are involved in the movement of subcellular components. (A) Schematic representation of Mcp6. The locations of the two coiled-coil motifs and the NLSs are indicated [as identified by PSORT II (http://psort.nibb.ac.jp/)]. (B) Schematic representation of Mcp6-related proteins of other species. The locations of homologous domains in these proteins are denoted by vertical white, grey and black arrowheads. Multiple sequence alignment shows homologous domains between Mcp6 from S. pombe (denoted Sp) and the related proteins from other species. Gaps inserted in the alignment to attain maximal homology are indicated by hyphens. The amino acids that are identical or similar among four or more of the seven species examined are shaded in black or grey. Abbreviations: MHC, myosin heavy chain; SMC, structural maintenance of chromosomes; Sp, Schizosaccharomyces pombe; At, Arabidopsis thaliana; Ce, Caenorhabditis elegans; Sc, Saccharomyces cerevisiae; Mm, Mus musculus; Xl, Xenopus laevis; Hs, Homo sapiens. The analysis of DNA sequences was performed by using the GENETYX program (Software Development, Tokyo, Japan). (C) Northern blot analysis of mcp6⁺ and aro3⁺ (loading control). Total RNA was extracted from CD16-1 (h⁺/h^-) and CD16-5 (h^-/h^-) cells at the indicated times after the induction of meiosis by nitrogen starvation. RNA was blotted and probed with the ORFs of mcp6⁺ and aro3⁺. The graph below indicates the meiotic profiles of the cells used for RNA extraction. The progression of meiosis was monitored every 2 hours after nitrogen starvation. The numbers of cells that bear one, two, three or four nuclei were assessed by counting the nuclei stained with Hoechst 33342. At least 200 cells were counted under the microscope. (D) Western blot analysis of the production of Mcp6-GFP and Meu13 (meiotic timing control) proteins during the synchronous meiosis of strain ST194. Tubulin levels were also examined as a loading control.

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