Ect2 and MgcRacGAP regulate the activation and function of Cdc42 in mitosis.

Ect2 and MgcRacGAP regulate the activation and function of Cdc42 in mitosis.
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ECT2和MGCRACGAP调节Cdc42在有丝分裂中的激活和功能。

DOI:
10.1083/jcb.200408085
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发表时间:
2005-01-17
影响因子:
7.8
通讯作者:
Narumiya, S
Narumiya, S
中科院分区:
生物学1区
文献类型:
--
作者:
Oceguera-Yanez, F;Kimura, K;Yasuda, S;Higashida, C;Kitamura, T;Hiraoka, Y;Haraguchi, T;Narumiya, S

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虽然Rho调节胞质分裂,但Cdc 42和Rac在有丝分裂中的功能知之甚少。我们最近提出Cdc 42通过调节纺锤体微管与动粒的双向附着在中期发挥作用。我们现在通过RNA干扰证实Cdc 42的作用,并确定Cdc 42激活和下调的机制。使用下拉分析,我们发现GTP-Cdc 42的水平在中期升高,而GTP-Rac的水平在有丝分裂中没有显著变化。Ect 2和MgcRacGAP的显性负突变体的过表达,分别是Rho GT3鸟嘌呤核苷酸交换因子和GT3激活蛋白,或通过RNA干扰耗尽Ect 2抑制有丝分裂中GTP-Cdc 42的这种变化。Ect 2的耗尽也损害微管附着到动粒,并导致前中期延迟和异常染色体分离,Cdc 42的耗尽或Ect 2和MgcRacGAP突变体的表达也是如此。这些结果表明,Ect 2和MgcRacGAP调节Cdc 42在有丝分裂中的激活和功能。
Although Rho regulates cytokinesis, little was known about the functions in mitosis of Cdc42 and Rac. We recently suggested that Cdc42 works in metaphase by regulating bi-orient attachment of spindle microtubules to kinetochores. We now confirm the role of Cdc42 by RNA interference and identify the mechanisms for activation and down-regulation of Cdc42. Using a pull-down assay, we found that the level of GTP-Cdc42 elevates in metaphase, whereas the level of GTP-Rac does not change significantly in mitosis. Overexpression of dominant-negative mutants of Ect2 and MgcRacGAP, a Rho GTPase guanine nucleotide exchange factor and GTPase activating protein, respectively, or depletion of Ect2 by RNA interference suppresses this change of GTP-Cdc42 in mitosis. Depletion of Ect2 also impairs microtubule attachment to kinetochores and causes prometaphase delay and abnormal chromosomal segregation, as does depletion of Cdc42 or expression of the Ect2 and MgcRacGAP mutants. These results suggest that Ect2 and MgcRacGAP regulate the activation and function of Cdc42 in mitosis.
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