Kurahashi Hiroshi, Imai Yoshiyuki, Yamamoto Masayuki
Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Hongo, Tokyo 113-0033, Japan.
Genes Cells. 2002 Apr;7(4):375-84. doi: 10.1046/j.1365-2443.2002.00526.x.
Tropomyosin is an actin-binding protein, which is thought to stabilize actin filaments and influence many aspects of F-actin. In fission yeast, the cdc8 gene encodes tropomyosin, and the gene product Cdc8p is known to be essential for the formation of the F-actin contractile ring and hence for cytokinesis in the mitotic cell cycle.
We isolated fission yeast mutants that were defective in cell fusion during conjugation. One of them turned out to carry a point mutation in cdc8. We found that the original temperature-sensitive cdc8 mutant frequently failed to undergo cell fusion when mated at a semi-permissive temperature. Additional cdc8 mutants isolated by targeted mutagenesis also showed defects in both cell fusion and cytokinesis. A decrease in the amount of intracellular Cdc8p also affected both, but cell growth was more severely blocked than cell fusion in this case. Immunostaining revealed that Cdc8p was localized as a spot at the cell-to-cell attachment site during conjugation, without overlapping with F-actin patches.
Tropomyosin Cdc8p is indispensable for cell fusion during conjugation in fission yeast. However, cell fusion appears to require fewer tropomyosin molecules than cytokinesis. We speculate that tropomyosin may organize a small F-actin-containing organelle at the cell-to-cell contact site in each mating cell, which plays a key role in cell fusion.
原肌球蛋白是一种肌动蛋白结合蛋白,被认为可稳定肌动蛋白丝并影响F-肌动蛋白的多个方面。在裂殖酵母中,cdc8基因编码原肌球蛋白,已知基因产物Cdc8p对于F-肌动蛋白收缩环的形成至关重要,因此对于有丝分裂细胞周期中的胞质分裂也至关重要。
我们分离出了在接合过程中细胞融合存在缺陷的裂殖酵母突变体。其中一个突变体在cdc8基因中携带一个点突变。我们发现,原来的温度敏感型cdc8突变体在半允许温度下交配时经常无法进行细胞融合。通过定向诱变分离出的其他cdc8突变体在细胞融合和胞质分裂方面也表现出缺陷。细胞内Cdc8p量的减少也对两者都有影响,但在这种情况下,细胞生长比细胞融合受到的阻碍更严重。免疫染色显示,在接合过程中,Cdc8p定位为细胞间附着位点处的一个斑点,不与F-肌动蛋白斑块重叠。
原肌球蛋白Cdc8p对于裂殖酵母接合过程中的细胞融合是不可或缺的。然而,细胞融合似乎比胞质分裂需要更少的原肌球蛋白分子。我们推测,原肌球蛋白可能在每个交配细胞的细胞间接触位点组织一个含少量F-肌动蛋白的细胞器,该细胞器在细胞融合中起关键作用。