Cantwell Helena, Nguyen Hieu, Kettenbach Arminja N, Heald Rebecca
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.
Department of Biochemistry and Cell Biology, Geisel School of Medicine at Dartmouth College, Hanover, NH, USA.
J Cell Biol. 2025 Aug 4;224(8). doi: 10.1083/jcb.202407154. Epub 2025 Jul 1.
The transition from meiotic divisions in the oocyte to embryonic mitoses is a critical step in animal development. Despite negligible changes to cell size and shape, following fertilization the small, barrel-shaped meiotic spindle is replaced by a large zygotic spindle that nucleates abundant astral microtubules at spindle poles. To probe underlying mechanisms, we applied a drug treatment approach using Ciona eggs and found that inhibition of casein kinase 2 (CK2) caused a shift from meiotic to mitotic-like spindle morphology with nucleation of robust astral microtubules, an effect reproduced in Xenopus egg cytoplasmic extracts. In both species, CK2 activity decreased at fertilization. Phosphoproteomic differences between Xenopus meiotic and mitotic extracts that also accompanied CK2 inhibition pointed to RanGTP-regulated factors as potential targets. Interfering with RanGTP-driven microtubule formation suppressed astral microtubule growth caused by CK2 inhibition. These data support a model in which CK2 activity attenuation at fertilization leads to activation of RanGTP-regulated microtubule effectors, inducing mitotic spindle morphology.
从卵母细胞减数分裂到胚胎有丝分裂的转变是动物发育中的关键步骤。尽管受精后细胞大小和形状变化可忽略不计,但小的桶状减数分裂纺锤体被一个大的合子纺锤体所取代,该合子纺锤体在纺锤体极处形成大量星体微管。为探究潜在机制,我们对海鞘卵采用药物处理方法,发现抑制酪蛋白激酶2(CK2)会导致纺锤体形态从减数分裂样转变为有丝分裂样,并形成强大的星体微管,这种效应在非洲爪蟾卵细胞质提取物中也能重现。在这两个物种中,受精时CK2活性均降低。非洲爪蟾减数分裂和有丝分裂提取物之间的磷酸化蛋白质组差异(同样伴随着CK2抑制)表明,RanGTP调节因子是潜在靶点。干扰RanGTP驱动的微管形成可抑制由CK2抑制引起的星体微管生长。这些数据支持了一个模型,即受精时CK2活性的减弱会导致RanGTP调节的微管效应器激活,从而诱导有丝分裂纺锤体形态。