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Fig. 1. Mug27 is a meiosis-specific serine/threonine protein kinase that is conserved in various species. (A) Schematic representation of Mug27 and of other members of the NDR family of proteins. The predicted protein-kinase domains (black box) are indicated. The nuclear-localization signal of Mug27 is also indicated (gray box). These motifs were identified by PSORT II (http://psort.nibb.ac.jp/) and Pfam (http://www.sanger.ac.uk/Software/Pfam). Hs, Homo sapien; Sc, Saccharomyces cerevisiae; Sp, Schizosaccharomyces pombe; Dm, Drosophila melanogaster; Ce, Caenorhabditis elegans. (B) The phylogenetic tree of Mug27. The relationships between the orthologous proteins were inferred by the neighbor-joining method. The numbers represent the phylogenetic distance. Both the sequence and phylogenetic analyses were performed using a GENETYX program (Software Development Co., Ltd). Tb, Trypanosoma brucei. (C) Western blot analysis of the production of Mug27-9Myc and Meu13 (meiotic timing control) proteins during the synchronous meiosis of strain SOP015. Tubulin levels were also examined as a loading control. (D) Northern blot analysis of mug27+, sid2+ and leu1+ (loading control) expression. Total RNA was extracted from CD16-1 (h+/h–) and CD16-5 (h–/h–) cells at the indicated times after nitrogen starvation. The former but not the latter strain enters meiosis upon nitrogen starvation. The RNA was blotted and probed with the ORFs of mug27+ and leu1+. The graphs indicates the meiotic profiles of the cells used for RNA extraction. The progression of meiosis was monitored every 2 hours after nitrogen starvation. The cells with one, two, three or four nuclei were enumerated by counting the Hoechst-33342-stained nuclei. At least 200 cells were counted under the microscope.
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