Cytoplasmic dynein plays a role in mammalian mitotic spindle formation.
Cytoplasmic dynein plays a role in mammalian mitotic spindle formation.
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DOI：
10.1083/jcb.123.4.849
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发表时间：
1993-11
影响因子：
7.8
通讯作者：
McIntosh, J R
中科院分区：
文献类型：
--
作者：
Vaisberg, E A;Koonce, M P;McIntosh, J R
有丝分裂纺锤体的形成和功能不仅取决于微管的组装/拆卸，而且还取决于马达酶的作用。细胞质动力蛋白定位于纺锤体，但它是否或如何在有丝分裂过程中发挥作用尚不清楚。我们克隆并表达了编码HeLa细胞和网骨藻细胞质动力蛋白重链的ATP水解位点的DNA片段。单特异性抗体已经产生的多肽，这些抑制动力蛋白的运动活性在体外。将它们注射到有丝分裂的哺乳动物细胞中，在有丝分裂的后期阶段，在前期或从诺考达唑治疗中恢复期间阻断纺锤体的形成。细胞变得与未分离的中心体和形式单极纺锤体。注射的抗体对染色体附着在双极纺锤体上或对后期的运动没有可检测到的影响。这些数据表明，细胞质动力蛋白在双极纺锤体形成的初始事件中起着独特而重要的作用，而它可能发挥的任何后续作用都是多余的。动力蛋白参与有丝分裂的可能机制进行了讨论。
The formation and functioning of a mitotic spindle depends not only on the assembly/disassembly of microtubules but also on the action of motor enzymes. Cytoplasmic dynein has been localized to spindles, but whether or how it functions in mitotic processes is not yet known. We have cloned and expressed DNA fragments that encode the putative ATP- hydrolytic sites of the cytoplasmic dynein heavy chain from HeLa cells and from Dictyostelium. Monospecific antibodies have been raised to the resulting polypeptides, and these inhibit dynein motor activity in vitro. Their injection into mitotic mammalian cells blocks the formation of spindles in prophase or during recovery from nocodazole treatment at later stages of mitosis. Cells become arrested with unseparated centrosomes and form monopolar spindles. The injected antibodies have no detectable effect on chromosome attachment to a bipolar spindle or on motions during anaphase. These data suggest that cytoplasmic dynein plays a unique and important role in the initial events of bipolar spindle formation, while any later roles that it may play are redundant. Possible mechanisms of dynein's involvement in mitosis are discussed.