Saskova Adela, Solc Petr, Baran Vladimir, Kubelka Michal, Schultz Richard M, Motlik Jan
Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Libechov, Czech Republic.
Cell Cycle. 2008 Aug;7(15):2368-76. doi: 10.4161/cc.6361. Epub 2008 May 29.
Aurora kinase A (AURKA), which is a centrosome-localized serine/threonine kinase crucial for cell cycle control, is critically involved in centrosome maturation and spindle assembly in somatic cells. Active T288 phosphorylated AURKA localizes to the centrosome in the late G(2) and also spreads to the minus ends of mitotic spindle microtubules. AURKA activates centrosomal CDC25B and recruits cyclin B1 to centrosomes. We report here functions for AURKA in meiotic maturation of mouse oocytes, which is a model system to study the G(2) to M transition. Whereas AURKA is present throughout the entire GV-stage oocyte with a clear accumulation on microtubule organizing centers (MTOC), active AURKA becomes entirely localized to MTOCs shortly before germinal vesicle breakdown. In contrast to somatic cells in which active AURKA is present at the centrosomes and minus ends of microtubules, active AURKA is mainly located on MTOCs at metaphase I (MI) in oocytes. Inhibitor studies using Roscovitine (CDK1 inhibitor), LY-294002 (PI3K inhibitor) and SH-6 (PKB inhibitor) reveal that activation of AURKA localized on MTOCs is independent on PI3K-PKB and CDK1 signaling pathways and MOTC amplification is observed in roscovitine- and SH-6-treated oocytes that fail to undergo nuclear envelope breakdown. Moreover, microinjection of Aurka mRNA into GV-stage oocytes cultured in 3-isobutyl-1-methyl xanthine (IBMX)-containing medium to prevent maturation also results in MOTC amplification in the absence of CDK1 activation. Overexpression of AURKA also leads to formation of an abnormal MI spindle, whereas RNAi-mediated reduction of AURKA interferes with resumption of meiosis and spindle assembly. Results of these experiments indicate that AURKA is a critical MTOC-associated component involved in resumption of meiosis, MTOC multiplication, proper spindle formation and the metaphase I-metaphase II transition.
极光激酶A(AURKA)是一种定位于中心体的丝氨酸/苏氨酸激酶,对细胞周期调控至关重要,在体细胞的中心体成熟和纺锤体组装中起着关键作用。活性T288磷酸化的AURKA在G2晚期定位于中心体,并且还扩散到有丝分裂纺锤体微管的负端。AURKA激活中心体的CDC25B并将细胞周期蛋白B1招募到中心体。我们在此报告AURKA在小鼠卵母细胞减数分裂成熟中的功能,小鼠卵母细胞是研究G2到M期转换的模型系统。虽然AURKA在整个GV期卵母细胞中均有存在,并在微管组织中心(MTOC)上明显积聚,但活性AURKA在生发泡破裂前不久完全定位于MTOC。与活性AURKA存在于中心体和微管负端的体细胞不同,活性AURKA在卵母细胞减数第一次分裂中期(MI)主要位于MTOC上。使用Roscovitine(CDK1抑制剂)、LY-294002(PI3K抑制剂)和SH-6(PKB抑制剂)进行的抑制剂研究表明,定位于MTOC的AURKA的激活独立于PI3K-PKB和CDK1信号通路,并且在未能经历核膜破裂的经roscovitine和SH-6处理的卵母细胞中观察到MOTC扩增。此外,将Aurka mRNA显微注射到在含3-异丁基-1-甲基黄嘌呤(IBMX)的培养基中培养以防止成熟的GV期卵母细胞中,在没有CDK1激活的情况下也会导致MOTC扩增。AURKA的过表达还会导致异常的MI纺锤体形成,而RNAi介导的AURKA减少会干扰减数分裂的恢复和纺锤体组装。这些实验结果表明,AURKA是参与减数分裂恢复、MTOC增殖、正确纺锤体形成以及减数第一次分裂中期到减数第二次分裂中期转换的关键MTOC相关成分。