Cimini Daniela, Wan Xiaohu, Hirel Christophe B, Salmon E D
Department of Biology, 607 Fordham Hall, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
Curr Biol. 2006 Sep 5;16(17):1711-8. doi: 10.1016/j.cub.2006.07.022.
Merotelic kinetochore orientation is a misattachment in which a single kinetochore binds microtubules from both spindle poles rather than just one and can produce anaphase lagging chromosomes, a major source of aneuploidy. Merotelic kinetochore orientation occurs frequently in early mitosis, does not block chromosome alignment at the metaphase plate, and is not detected by the spindle checkpoint. However, microtubules to the incorrect pole are usually significantly reduced or eliminated before anaphase. We discovered that the frequency of lagging chromosomes in anaphase is very sensitive to partial inhibition of Aurora kinase activity by ZM447439 at a dose, 3 microM, that has little effect on histone phosphorylation, metaphase chromosome alignment, and cytokinesis in PtK1 cells. Partial Aurora kinase inhibition increased the frequency of merotelic kinetochores in late metaphase, and the fraction of microtubules to the incorrect pole. Measurements of fluorescence dissipation after photoactivation showed that kinetochore-microtubule turnover in prometaphase is substantially suppressed by partial Aurora kinase inhibition. Our results support a preanaphase correction mechanism for merotelic attachments in which correct plus-end attachments are pulled away from high concentrations of Aurora B at the inner centromere, and incorrect merotelic attachments are destabilized by being pulled toward the inner centromere.
着丝粒错向是一种错误连接,即单个着丝粒与来自纺锤体两极的微管结合,而不是仅与一极的微管结合,可产生后期滞后染色体,这是非整倍体的主要来源。着丝粒错向在有丝分裂早期频繁发生,不阻碍染色体在中期板上排列,且不能被纺锤体检查点检测到。然而,在后期之前,连接到错误极的微管通常会显著减少或消失。我们发现,在PtK1细胞中,用ZM447439以3微摩尔的剂量部分抑制极光激酶活性时,后期滞后染色体的频率对其非常敏感,而该剂量对组蛋白磷酸化、中期染色体排列和胞质分裂几乎没有影响。部分抑制极光激酶会增加中期后期着丝粒错向的频率以及连接到错误极的微管比例。光激活后荧光消散的测量结果表明,部分抑制极光激酶会显著抑制前中期着丝粒-微管的周转。我们的结果支持一种针对着丝粒错向的后期前校正机制,即正确的正端连接会从着丝粒内部高浓度的极光B处被拉开,而错误的着丝粒错向连接会因被拉向着丝粒内部而变得不稳定。