Meireles Ana M, Fisher Katherine H, Colombié Nathalie, Wakefield James G, Ohkura Hiroyuki
Wellcome Trust Centre for Cell Biology, The University of Edinburgh, Edinburgh, Scotland, UK.
J Cell Biol. 2009 Mar 23;184(6):777-84. doi: 10.1083/jcb.200811102. Epub 2009 Mar 16.
The bipolar spindle forms without centrosomes naturally in female meiosis and by experimental manipulation in mitosis. Augmin is a recently discovered protein complex required for centrosome-independent microtubule generation within the spindle in Drosophila melanogaster cultured cells. Five subunits of Augmin have been identified so far, but neither their organization within the complex nor their role in developing organisms is known. In this study, we report a new Augmin subunit, wee Augmin component (Wac). Wac directly interacts with another Augmin subunit, Dgt2, via its coiled-coil domain. Wac depletion in cultured cells, especially without functional centrosomes, causes severe defects in spindle assembly. We found that a wac deletion mutant is viable but female sterile and shows only a mild impact on somatic mitosis. Unexpectedly, mutant female meiosis showed robust microtubule assembly of the acentrosomal spindle but frequent chromosome misalignment. For the first time, this study establishes the role of an Augmin subunit in developing organisms and provides an insight into the architecture of the complex.
双极纺锤体在雌性减数分裂过程中自然形成且无需中心体,在有丝分裂中则通过实验操作形成。Augmin是最近发现的一种蛋白质复合物,在果蝇培养细胞的纺锤体内,它是独立于中心体产生微管所必需的。到目前为止,已鉴定出Augmin的五个亚基,但它们在复合物中的组织方式以及在发育生物体中的作用尚不清楚。在本研究中,我们报告了一种新的Augmin亚基,即微小Augmin组分(Wac)。Wac通过其卷曲螺旋结构域直接与另一个Augmin亚基Dgt2相互作用。在培养细胞中耗尽Wac,尤其是在没有功能性中心体的情况下,会导致纺锤体组装出现严重缺陷。我们发现,wac缺失突变体是可存活的,但雌性不育,并且对体细胞有丝分裂仅表现出轻微影响。出乎意料的是,突变体雌性减数分裂显示无中心体纺锤体的微管组装强劲,但染色体频繁排列错误。本研究首次确定了Augmin亚基在发育生物体中的作用,并深入了解了该复合物的结构。