Yamashita Akira, Sato Masamitsu, Fujita Akiko, Yamamoto Masayuki, Toda Takashi
Molecular Genetics Research Laboratory, Graduate School of Science, University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Mol Biol Cell. 2005 Mar;16(3):1378-95. doi: 10.1091/mbc.e04-10-0859. Epub 2005 Jan 12.
The Ase1/Prc1 proteins constitute a conserved microtubule-associated protein family that is implicated in central spindle formation and cytokinesis. Here we characterize a role for fission yeast Ase1. Ase1 localizes to microtubule overlapping zones and displays dynamic alterations of localization during the cell cycle. In particular, its spindle localization during metaphase is reduced substantially, followed by robust appearance at the spindle midzone in anaphase. ase1 deletions are viable but defective in nuclear and septum positioning and completion of cytokinesis, which leads to diploidization and chromosome loss. Time-lapse imaging shows that elongating spindles collapse abruptly in the middle of anaphase B. Either absence or overproduction of Ase1 results in profound defects on microtubule bundling in an opposed manner, indicating that Ase1 is a dose-dependent microtubule-bundling factor. In contrast microtubule nucleating activities are not noticeably compromised in ase1 mutants. During meiosis astral microtubules are not bundled and oscillatory nuclear movement is impaired significantly. The Aurora kinase does not correctly localize to central spindles in the absence of Ase1. Finally Ase1 acts as a regulatory component in the cytokinesis checkpoint that operates to inhibit nuclear division when the cytokinesis apparatus is perturbed. Ase1, therefore, couples anaphase completion with cytokinesis upon cell division.
Ase1/Prc1蛋白构成了一个保守的微管相关蛋白家族,该家族与中央纺锤体的形成及胞质分裂有关。在此,我们阐述了裂殖酵母Ase1的作用。Ase1定位于微管重叠区,并在细胞周期中呈现出定位的动态变化。特别地,其在中期的纺锤体定位显著降低,随后在后期纺锤体中间区大量出现。缺失ase1是可行的,但在细胞核和隔膜定位以及胞质分裂完成方面存在缺陷,这会导致二倍体化和染色体丢失。延时成像显示,在后期B的中期,伸长的纺锤体会突然塌陷。Ase1的缺失或过量表达都会以相反的方式导致微管束出现严重缺陷,这表明Ase1是一种剂量依赖性的微管束因子。相比之下,ase1突变体中的微管成核活性并未受到明显影响。在减数分裂过程中,星体微管不会成束,振荡核运动也会受到显著损害。在没有Ase1的情况下,极光激酶不能正确定位于中央纺锤体。最后,Ase1在胞质分裂检查点中作为一个调节成分发挥作用,当胞质分裂装置受到干扰时,该检查点会抑制核分裂。因此,Ase1在细胞分裂时将后期完成与胞质分裂联系起来。