Spatial organization of cytokinesis signaling reconstituted in a cell-free system.

Spatial organization of cytokinesis signaling reconstituted in a cell-free system.
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DOI:
10.1126/science.1256773
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发表时间:
2014-10-10
期刊:
Science (New York, N.Y.)
影响因子:
--
通讯作者:
Mitchison TJ
Mitchison TJ
中科院分区:
其他
文献类型:
--
作者:
Nguyen PA;Groen AC;Loose M;Ishihara K;Wühr M;Field CM;Mitchison TJ

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在动物细胞分裂过程中,卵裂沟由微管定位,微管向细胞中平面的肌动蛋白皮层发出信号。我们开发了一种无细胞系统,利用非洲爪蟾卵的细胞质提取物来重现胞质分裂信号。微管从人工中心体中长成紫苑,并相遇形成反平行重叠区域。这些区域阻止了邻近紫苑的相互渗透,并招募了胞质分裂中区蛋白,包括染色体乘客复合体(CPC)和中枢蛋白。 CPC 被运输到重叠区域,这需要两种运动蛋白 Kif4A 和 Kif20A 旁系同源蛋白。使用支持的脂质双层来模拟质膜,我们观察到在微管重叠处招募了裂解沟标记物,包括 RhoA-GTP 报告基因。该系统为理解胞质分裂信号的生物物理学开辟了进一步的途径。
During animal cell division, the cleavage furrow is positioned by microtubules that signal to the actin cortex at the cell midplane. We developed a cell-free system to recapitulate cytokinesis signaling using cytoplasmic extract from Xenopus eggs. Microtubules grew out as asters from artificial centrosomes and met to organize antiparallel overlap zones. These zones blocked interpenetration of neighboring asters and recruited cytokinesis midzone proteins including the Chromosomal Passenger Complex (CPC) and Centralspindlin. The CPC was transported to overlap zones, which required two motor proteins, Kif4A and a Kif20A paralog. Using supported lipid bilayers to mimic the plasma membrane, we observed recruitment of cleavage furrow markers, including a RhoA-GTP reporter, at microtubule overlaps. This system opens further approaches to understanding the biophysics of cytokinesis signaling.
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