Lippincott J, Li R
Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
J Cell Biol. 1998 Dec 28;143(7):1947-60. doi: 10.1083/jcb.143.7.1947.
We previously showed that the budding yeast Saccharomyces cerevisiae assembles an actomyosin-based ring that undergoes a contraction-like size change during cytokinesis. To learn more about the biochemical composition and activity of this ring, we have characterized the in vivo distribution and function of Cyk2p, a budding yeast protein that exhibits significant sequence similarity to the cdc15/PSTPIP family of cleavage furrow proteins. Video microscopy of cells expressing green fluorescent protein (GFP)-tagged Cyk2p revealed that Cyk2p forms a double ring that coincides with the septins through most of the cell cycle. During cytokinesis, however, the Cyk2 double ring merges with the actomyosin ring and exhibits a contraction-like size change that is dependent on Myo1p. The septin double ring, in contrast, does not undergo the contraction-like size change but the separation between the two rings increases during cytokinesis. These observations suggest that the septin-containing ring is dynamically distinct from the actomyosin ring and that Cyk2p transits between the two types of structures. Gene disruption of CYK2 does not affect the assembly of the actomyosin ring but results in rapid disassembly of the ring during the contraction phase, leading to incomplete cytokinesis, suggesting that Cyk2p has an important function in modulating the stability of the actomyosin ring during contraction. Overexpression of Cyk2p also blocks cytokinesis, most likely due to a loss of the septins from the bud neck, indicating that Cyk2p may also play a role in regulating the localization of the septins.
我们之前发现,出芽酵母酿酒酵母会组装一个基于肌动球蛋白的环,该环在胞质分裂过程中会经历类似收缩的大小变化。为了更深入了解这个环的生化组成和活性,我们对Cyk2p在体内的分布和功能进行了表征,Cyk2p是一种出芽酵母蛋白,与分裂沟蛋白的cdc15/PSTPIP家族具有显著的序列相似性。对表达绿色荧光蛋白(GFP)标记的Cyk2p的细胞进行视频显微镜观察发现,Cyk2p形成一个双环,在细胞周期的大部分时间里与隔膜蛋白重合。然而,在胞质分裂期间,Cyk2双环与肌动球蛋白环合并,并呈现出类似收缩的大小变化,这种变化依赖于Myo1p。相比之下,隔膜蛋白双环不会经历类似收缩的大小变化,但在胞质分裂期间两个环之间的间距会增加。这些观察结果表明,含隔膜蛋白的环与肌动球蛋白环在动态上是不同的,并且Cyk2p在这两种结构之间转变。CYK2基因的破坏不会影响肌动球蛋白环的组装,但会导致该环在收缩阶段迅速解体,从而导致不完全胞质分裂,这表明Cyk2p在调节收缩过程中肌动球蛋白环的稳定性方面具有重要作用。Cyk2p的过表达也会阻断胞质分裂,最有可能是由于芽颈处隔膜蛋白的缺失,这表明Cyk2p可能也在调节隔膜蛋白的定位中发挥作用。