Shannon Katie B, Canman Julie C, Ben Moree C, Tirnauer Jennifer S, Salmon E D
Department of Biology, University of North Carolina, Chapel Hill, NC 27599-3280, USA.
Mol Biol Cell. 2005 Sep;16(9):4423-36. doi: 10.1091/mbc.e04-11-0974. Epub 2005 Jun 22.
How microtubules act to position the plane of cell division during cytokinesis is a topic of much debate. Recently, we showed that a subpopulation of stable microtubules extends past chromosomes and interacts with the cell cortex at the site of furrowing, suggesting that these stabilized microtubules may stimulate contractility. To test the hypothesis that stable microtubules can position furrows, we used taxol to rapidly suppress microtubule dynamics during various stages of mitosis in PtK1 cells. Cells with stabilized prometaphase or metaphase microtubule arrays were able to initiate furrowing when induced into anaphase by inhibition of the spindle checkpoint. In these cells, few microtubules contacted the cortex. Furrows formed later than usual, were often aberrant, and did not progress to completion. Images showed that furrowing correlated with the presence of one or a few stable spindle microtubule plus ends at the cortex. Actin, myosin II, and anillin were all concentrated in these furrows, demonstrating that components of the contractile ring can be localized by stable microtubules. Inner centromere protein (INCENP) was not found in these ingressions, confirming that INCENP is dispensable for furrow positioning. Taxol-stabilization of the numerous microtubule-cortex interactions after anaphase onset delayed furrow initiation but did not perturb furrow positioning. We conclude that taxol-stabilized microtubules can act to position the furrow and that loss of microtubule dynamics delays the timing of furrow onset and prevents completion. We discuss our findings relative to models for cleavage stimulation.
在胞质分裂过程中微管如何作用来确定细胞分裂平面是一个备受争议的话题。最近,我们发现一群稳定的微管延伸至染色体之外,并在缢缩部位与细胞皮层相互作用,这表明这些稳定的微管可能刺激收缩性。为了验证稳定的微管能够确定缢缩位置这一假说,我们使用紫杉醇在PtK1细胞有丝分裂的各个阶段快速抑制微管动力学。当通过抑制纺锤体检查点诱导进入后期时,前期或中期微管阵列稳定的细胞能够启动缢缩。在这些细胞中,很少有微管接触皮层。缢缩形成比正常情况晚,常常异常,且无法进展至完成。图像显示缢缩与皮层处一根或几根稳定的纺锤体微管正端的存在相关。肌动蛋白、肌球蛋白II和锚蛋白都集中在这些缢缩处,表明收缩环的成分可由稳定的微管定位。在这些缢缩处未发现着丝粒内蛋白(INCENP),证实INCENP对于缢缩定位并非必需。后期开始后大量微管与皮层相互作用经紫杉醇稳定化会延迟缢缩起始,但不会干扰缢缩定位。我们得出结论,紫杉醇稳定的微管能够作用于确定缢缩位置,微管动力学的丧失会延迟缢缩起始时间并阻止其完成。我们结合分裂刺激模型讨论了我们的发现。