有丝分裂激酶Nek2调节分裂期中心体结构及功能的机制研究
结题报告
批准号:
31970673
项目类别:
面上项目
资助金额:
58.0 万元
负责人:
余娴文
依托单位:
学科分类:
细胞增殖及细胞周期
结题年份:
2023
批准年份:
2019
项目状态:
已结题
项目参与者:
余娴文
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中文摘要
我们发表的文章揭示了中心体蛋白Cep85通过调控Nek2激酶活性在时空上确保了中心体在间期不过早分离而在分裂早期适时分离的功能。但是反过来Nek2是否调控Cep85功能至今没有被报道。我们的前期研究发现:Cep85在细胞内形成多聚体,影响分裂期中心体的微管组织能力;Nek2能够磷酸化Cep85,激酶活性是Cep85形成多聚体所必需;Nek2敲低减弱分裂期中心体的Cep85及微管结合力。因此,我们推测Nek2可能是通过调节多聚体的形成增强Cep85在中心体上的聚集来参与调节分裂期中心体的结构和功能。我们将在构建稳转细胞系及鉴定Cep85磷酸化位点的基础上,从内源及胚胎发育水平上首先研究Cep85形成多聚体及聚集到分裂期中心体上的调节机制,再研究Nek2只能在细胞内磷酸化Cep85的机制,彻底阐明Nek2在分裂早期通过修饰Cep85来提高它在中心体上的聚集、参与调节中心体结构和功能这一新途径。
英文摘要
Due to the lack of mechanistic study, it is for a long time unknown whether Nek2 kinase is indeed involved in the regulation of the structure and function of centrosomes in mitotic stage. Recently, we have demonstrated in our research article (JCS, 2015) that Cep85 is a negative regulator for Nek2A to prevent the precocious separation of centrosomes in interphase by binding to and suppressing Nek2A, a mitotic kinase that is responsible for initiating centrosome separation at G2/M. We have further demonstrated in our newly published paper (iScience, 2019) that Cep85-assisting Nek2A activation by Plk1 at G2/M is a critical step for Nek2A to achieve a threshold activity to initiate centrosome separation at the onset of mitosis. We have also revealed that Cep85 greatly increases its accumulation at centrosomes in mitotic stage and its depletion results in the significantly reduction of γ-tubulin on mitotic centrosomes. However, it remained unknown the underlying mechanisms regulation the accumulation of Cep85 to the mitotic centrosomes..Our preliminary data show that Cep85 depletion results in the significant reduction of γ-tubulin on centrosomes and the spindle formation in mitotic stage., and Cep85 itself can form an oligomer in cells. Once co-expressed, wildtype but not the kinase-dead mutant K37R of Nek2A can facilitate the formation of Cep85 oligomer. Similar to Nek2A, Nek2B can also phosphorylate Cep85 in cells and enhance the formation of Cep85 oligomer. Nek2A depletion significantly reduces the centrosomal accumulation of Cep85 and γ-tubulin, suggesting that Nek2 is involved in the regulation of the function of mitotic centrosomes through controlling Cep85 accumulation at centrosomes. We therefore will further investigate the mechanisms regulating the oligomer formation and the centrosomal accumulation of Cep85 in cells and in zebrafish embryos to reveal of the roles of Nek2 playing in the regulation of the structure organization and function of mitotic centrosomes.
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中心体结合蛋白Cep88在中心体复制及斑马鱼早期胚胎发育中的功能研究
  • 批准号:
    31471260
  • 项目类别:
    面上项目
  • 资助金额:
    80.0万元
  • 批准年份:
    2014
  • 负责人:
    余娴文
  • 依托单位:
调控动纤毛形成与功能的分子机制研究
  • 批准号:
    31171286
  • 项目类别:
    面上项目
  • 资助金额:
    65.0万元
  • 批准年份:
    2011
  • 负责人:
    余娴文
  • 依托单位:
国内基金
海外基金