Live-cell analysis of mitotic spindle formation in taxol-treated cells.
Live-cell analysis of mitotic spindle formation in taxol-treated cells.
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DOI：
10.1002/cm.20283
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发表时间：
2008-08
影响因子：
--
通讯作者：
Hinchcliffe, Edward H.
中科院分区：
文献类型：
--
作者：
Hornick, Jessica E.;Bader, Jason R.;Tribble, Emily K.;Trimble, Kayleigh;Breunig, J. Scott;Halpin, Elizabeth S.;Vaughan, Kevin T.;Hinchcliffe, Edward H.
紫杉醇抑制单个微管的动态行为，诱导多极有丝分裂纺锤体。然而，关于紫杉醇在体内破坏正常双极纺锤体组装的机制知之甚少。利用表达GFP-α微管蛋白的细胞的实时成像，我们检测了紫杉醇处理后的纺锤体组装。我们发现，当紫杉醇处理的细胞进入有丝分裂时，微管网络从中心体到细胞皮层发生了戏剧性的重新分配。当它们排列在那里时，皮层微管将NuMA招募到它们嵌入的末端，然后是运动蛋白运动HSET。这些皮层微管随后发芽形成胞丝细胞，它们融合成多极纺锤体。细胞质动力蛋白和动力蛋白不会重新定位到皮质微管，p50“动力蛋白”的过度表达破坏动力蛋白/动力蛋白的相互作用并不能阻止胞浆的形成。在纺锤极开始形成之前加入紫杉醇会导致多极，但在新生的纺锤极可见之后（但在NEB完全形成之前）加入紫杉醇会导致双极纺锤。我们的研究结果表明，紫杉醇可以阻止关键成分（如NuMA）快速运输到新生的纺锤极。最终的结果是有丝分裂纺锤极凝聚力的丧失，微管的重新分布和胞浆的形成。
Taxol functions to suppress the dynamic behavior of individual microtubules, and induces multipolar mitotic spindles. However, little is known about the mechanisms by which taxol disrupts normal bipolar spindle assembly in vivo. Using live imaging of GFP-α tubulin expressing cells, we examined spindle assembly after taxol treatment. We find that as taxol-treated cells enter mitosis, there is a dramatic redistribution of the microtubule network from the centrosomes to the cell cortex. As they align there, the cortical microtubules recruit NuMA to their embedded ends, followed by the kinesin motor HSET. These cortical microtubules then bud off to form cytasters, which fuse into multipolar spindles. Cytoplasmic dynein and dynactin do not re-localize to cortical microtubules, and disruption of dynein/dynactin interactions by over-expression of p50 “dynamitin” does not prevent cytaster formation. Taxol added well before spindle poles begin to form induces multipolarity, but taxol added after nascent spindle poles are visible—but before NEB is complete—results in bipolar spindles. Our results suggest that taxol prevents rapid transport of key components, such as NuMA, to the nascent spindle poles. The net result is loss of mitotic spindle pole cohesion, microtubule re-distribution, and cytaster formation.
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DOI：
10.1187/cbe.05-02-0064
发表时间：
2005-01-01
期刊：
Cell biology education
影响因子：
--
作者：
Hinchcliffe, Edward H
通讯作者：
Hinchcliffe, Edward H
影响因子：
3.3
作者：
Piehl, M;Cassimeris, L
通讯作者：
Cassimeris, L
DOI：
10.1083/jcb.141.1.163
发表时间：
1998-04-06
期刊：
The Journal of cell biology
影响因子：
--
作者：
Dictenberg JB;Zimmerman W;Sparks CA;Young A;Vidair C;Zheng Y;Carrington W;Fay FS;Doxsey SJ
通讯作者：
Doxsey SJ
影响因子：
21.3
作者：
Giannakakou, P;Sackett, DL;Fojo, T
通讯作者：
Fojo, T
DOI：
10.1083/jcb.135.2.399
发表时间：
1996-10
期刊：
The Journal of cell biology
影响因子：
--
作者：
Gaglio T;Saredi A;Bingham JB;Hasbani MJ;Gill SR;Schroer TA;Compton DA
通讯作者：
Compton DA