Lamb N J, Fernandez A, Watrin A, Labbé J C, Cavadore J C
Cell Biology, CRBM, CNRS/INSERM, Montpellier, France.
Cell. 1990 Jan 12;60(1):151-65. doi: 10.1016/0092-8674(90)90725-t.
We have examined the effects of elevating the intracellular levels of p34cdc2 kinase by microinjection into living mammalian cells. These studies reveal rapid and dramatic changes in cell shape with cells becoming round and losing the bulk of their cell-substratum contact. Such effects were induced at all times in the cell cycle except at S phase and were fully reversible at S phase or mitosis. Similar results were obtained with the homogeneous catalytic subunit of p34cdc2 kinase or p34cdc2 kinase associated with cyclin B. These alterations were accompanied by a marked reduction in interphase microtubules without the spindle formation, actin microfilament redistribution, and premature chromatin condensation. Although these changes closely mimic the events occurring during early phases of mitosis, p34cdc2 kinase-injected cells were not induced to pass further into division. These data provide detailed evidence that p34cdc2 kinase plays a major prerequisite role in the rearrangement of cellular structures associated with mammalian cell mitosis.
我们通过向活的哺乳动物细胞中显微注射来提高细胞内p34cdc2激酶的水平,并研究了其作用效果。这些研究揭示了细胞形状迅速且显著的变化,细胞变得圆钝,失去了大部分与底物的接触。除了在S期外,细胞周期的所有阶段都会诱导产生这种效应,并且在S期或有丝分裂期这种效应是完全可逆的。用p34cdc2激酶的同源催化亚基或与细胞周期蛋白B相关的p34cdc2激酶也得到了类似的结果。这些改变伴随着间期微管的显著减少,且没有纺锤体形成、肌动蛋白微丝重新分布以及染色质过早凝聚。尽管这些变化与有丝分裂早期发生的事件极为相似,但注射了p34cdc2激酶的细胞并没有被诱导进一步进入分裂阶段。这些数据提供了详细的证据,表明p34cdc2激酶在与哺乳动物细胞有丝分裂相关的细胞结构重排中起着主要的先决作用。