Yoshida Satoshi, Asakawa Kazuhide, Toh-e Akio
Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, 113-0033 Tokyo, Japan.
Curr Biol. 2002 Jun 4;12(11):944-50. doi: 10.1016/s0960-9822(02)00870-9.
Budding yeast Cdc14 phosphatase plays essential roles in mitotic exit. Cdc14 is sequestered in the nucleolus by its inhibitor Net1/Cfi1 and is only released from the nucleolus during anaphase to inactivate mitotic CDK. It is believed that the mitotic exit network (MEN) is required for the release of Cdc14 from the nucleolus because liberation of Cdc14 by net1/cfi1 mutations bypasses the essential role of the MEN. But how the MEN residing at the spindle pole body (SPB) controls the association of Cdc14 with Net1/Cfi1 in the nucleolus is not yet understood. We found that Cdc14-5GFP was released from the nucleolus in the MEN mutants (tem1, cdc15, dbf2, and nud1), but not in the cdc5 cells during early anaphase. The Cdc14 liberation from the nucleolus was inhibited by the Mad2 checkpoint and by the Bub2 checkpoint in a different manner when microtubule organization was disrupted. We observed Cdc14-5GFP at the SPB in addition to the nucleolus. The SPB localization of Cdc14 was significantly affected by the MEN mutations and the bub2 mutation. We conclude that Cdc14 is released from the nucleolus at the onset of anaphase in a CDC5-dependent manner and that MEN factors possibly regulate Cdc14 release from the SPB.
出芽酵母Cdc14磷酸酶在有丝分裂退出过程中发挥着至关重要的作用。Cdc14被其抑制剂Net1/Cfi1隔离在核仁中,仅在后期从核仁中释放出来以失活有丝分裂周期蛋白依赖性激酶(CDK)。据信,有丝分裂退出网络(MEN)是Cdc14从核仁中释放所必需的,因为net1/cfi1突变导致的Cdc14释放绕过了MEN的重要作用。但是,位于纺锤极体(SPB)的MEN如何控制Cdc14与核仁中Net1/Cfi1的结合尚不清楚。我们发现,在有丝分裂后期早期,Cdc14-5GFP在MEN突变体(tem1、cdc15、dbf2和nud1)中从核仁释放,但在cdc5细胞中未释放。当微管组织被破坏时,Mad2检查点和Bub2检查点以不同方式抑制了Cdc14从核仁的释放。除了核仁外,我们还在纺锤极体处观察到了Cdc14-5GFP。Cdc14在纺锤极体的定位受到MEN突变和bub2突变的显著影响。我们得出结论,Cdc14在后期开始时以依赖CDC5的方式从核仁释放,并且MEN因子可能调节Cdc14从纺锤极体的释放。