Wakefield J G, Huang J Y, Raff J W
Wellcome/CRC Institute and Department of Genetics, University of Cambridge, Cambridge, UK.
Curr Biol. 2000 Nov 2;10(21):1367-70. doi: 10.1016/s0960-9822(00)00776-4.
We reported previously that the disappearance of cyclin B at the end of mitosis in early Drosophila embryos starts at centrosomes and spreads into the spindle [1]. Here, we used a novel mutation, centrosome fall off (cfo), to investigate whether centrosomes are required to initiate the disappearance of cyclin B from the spindle. In embryos laid by homozygous cfo mutant mothers, the centrosomes co-ordinately detached from the mitotic spindle during mitosis, and the centrosomeless spindles arrested at anaphase. Cyclin B levels decreased on the detached centrosomes, but not on the arrested centrosomeless spindles, presumably explaining why the spindles arrest in anaphase in these embryos. We found that the expression of a non-degradable cyclin B in embryos also caused an anaphase arrest, but most centrosomes remained attached to the arrested spindles, and non-degradable cyclin B levels remained high on both the centrosomes and spindles. These findings suggest that the disappearance of cyclin B from centrosomes and spindles is closely linked to its destruction, and that a connection between centrosomes and spindles is required for the proper destruction of the spindle-associated cyclin B in early Drosophila embryos. These results may have important implications for the mechanism of the spindle-assembly checkpoint, as they suggest that unattached kinetochores may arrest cells in mitosis, at least in part, by signalling to centrosomes to block the initiation of cyclin B destruction.
我们之前报道过,在果蝇早期胚胎有丝分裂末期,细胞周期蛋白B的消失始于中心体,并扩散到纺锤体中[1]。在此,我们利用一种新的突变体——中心体脱落(cfo),来研究中心体是否是启动细胞周期蛋白B从纺锤体消失所必需的。在纯合cfo突变体母体所产的胚胎中,中心体在有丝分裂期间协调地从有丝分裂纺锤体上脱离,无中心体的纺锤体停滞在后期。细胞周期蛋白B在脱离的中心体上水平下降,但在停滞的无中心体纺锤体上则不然,这大概解释了为什么这些胚胎中的纺锤体在后期停滞。我们发现,在胚胎中表达不可降解的细胞周期蛋白B也会导致后期停滞,但大多数中心体仍附着在停滞的纺锤体上,不可降解的细胞周期蛋白B在中心体和纺锤体上的水平都保持较高。这些发现表明,细胞周期蛋白B从中心体和纺锤体的消失与其降解密切相关,并且在果蝇早期胚胎中,中心体与纺锤体之间的联系对于纺锤体相关细胞周期蛋白B的正常降解是必需的。这些结果可能对纺锤体组装检查点的机制具有重要意义,因为它们表明未附着的动粒可能至少部分地通过向中心体发出信号以阻止细胞周期蛋白B降解的启动,从而使细胞停滞在有丝分裂期。