Oakley B R, Morris N R
J Cell Biol. 1983 Apr;96(4):1155-8. doi: 10.1083/jcb.96.4.1155.
In order to develop a method for obtaining mitotic synchrony in aspergillus nidulans, we have characterized previously isolated heat-sensitive nim mutations that block the nuclear division cycle in interphase at restrictive temperature. After 3.5 h at restrictive temperature the mitotic index of a strain carrying one of these mutations, nimA5, was 0, but when this strain was subsequently shifted from restrictive to permissive temperature the mitotic index increased rapidly, reaching a maximum of 78 percent after 7.5 min. When this strain was examined electron-microscopically, mitotic spindles were absent at restrictive temperature. From these data we conclude that at restrictive temperature nimA5 blocks the nuclear division cycle at a point immediately preceding the initiation of chromosomal condensation and mitotic microtubule assembly, and upon shifting to permissive control over the initiation of microtubule assembly and chromosomal condensation in vivo through a simple temperature shift and, consequently, nimA5 should be a powerful tool for studying these processes. Electron-microscopic examination of spindles of material synchronized in this manner reveals that spindle formation, although very rapid, is gradual in the sense that spindle microtubule numbers increase as spindle formation proceeds.
为了开发一种在构巢曲霉中获得有丝分裂同步性的方法,我们对先前分离出的热敏感nim突变进行了表征,这些突变在限制温度下会在间期阻断核分裂周期。在限制温度下处理3.5小时后,携带其中一个突变(nimA5)的菌株的有丝分裂指数为0,但当该菌株随后从限制温度转移到允许温度时,有丝分裂指数迅速增加,在7.5分钟后达到最大值78%。当对该菌株进行电子显微镜检查时,在限制温度下没有发现有丝分裂纺锤体。从这些数据我们得出结论,在限制温度下,nimA5在染色体浓缩和有丝分裂微管组装开始之前的一个点阻断核分裂周期,并且在转移到允许温度后,通过简单的温度变化在体内控制微管组装和染色体浓缩的起始,因此,nimA5应该是研究这些过程的有力工具。对以这种方式同步化的材料的纺锤体进行电子显微镜检查发现,纺锤体形成虽然非常迅速,但从纺锤体微管数量随着纺锤体形成过程而增加的意义上来说是渐进的。