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Cyclin dependent kinase cascade.

细胞周期依赖性激酶级联。

基本信息

批准号：
13043057
负责人：
INAGAKI Masaki
金额：
$ 38.27万
依托单位：
Aichi Cancer Center (Research Institute)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2005
项目状态：
已结题

项目摘要

Mitotic chromosomal dynamics is regulated by the coordinated activities of many mitotic kinases, such as Cdc2 (Cdkl), Aurora-B or Polo-like kinase 1 (P1k1), but mechanisms of its coordination remain unknown. We demonstrated two novel Cdc2 kinase cascades. 1) We demonstrated a direct interaction between Plkl and vimentin-Ser55 phosphorylated by Cdc2, an event which led to Plkl activation and further vimentin phosphorylation by Plkl. In vitro analyses revealed that Plkl phosphorylated vimentin at,・1 mol phosphate/mol substrate, which partly inhibited its filament forming ability. Plkl phosphorylated Ser82 on vimentin in vitro, and this phosphorylation was elevated from metaphase and was maintained until the end of mitosis. This elevation thus followed the Cdc2-induced vimentin-Ser55 phosphorylation. Depletion of Plkl by RNA interference impaired the elevation of vimentin-Ser82 phosphorylation in mitosis. These results indicated a novel mechanism that Cdc2 regulated mitotic vimentin phosphorylation via not only a direct enzyme reaction but also Plkl recruitment to vimentin. 2) We found that Cdc2 phosphorylates Thr59 and Thr388 on inner centromere protein (INCENP), which regulates localization and kinase activity of Aurora-B, from prophase to metaphase. INCENP depletion disrupts Plkl localization specifically at the kinetochore. This phenotype is rescued by the exogenous expression of INCENP wild type (WT) and INCENP mutated at Thr59 to Ala (T59A), but not at Thr388 to Ala (T388A). The replacement of endogenous INCENP with T388A resulted in the delay of progression from metaphase to anaphage. Our findings suggested that INCENP phosphorylation by Cdc2 is necessary for the recruitment of Plkl to the kinetochore, and the complex formation of Plkl and Aurora-B on INCENP may play critical roles in the regulation of chromosomal dynamics.

有丝分裂染色体动力学受许多有丝分裂激酶的协调活动调节，如Cdc2 (Cdkl)， Aurora-B或polo样激酶1 (P1k1)，但其协调机制尚不清楚。我们证明了两个新的Cdc2激酶级联。1)我们证明了Plkl与Cdc2磷酸化的vimentin- ser55之间的直接相互作用，这一事件导致Plkl活化和Plkl进一步磷酸化vimentin。体外分析表明，Plkl以1 mol磷酸/mol底物磷酸化vimentin，部分抑制其成丝能力。Plkl在体外磷酸化vimentin上的Ser82，这种磷酸化从中期开始升高，并维持到有丝分裂结束。因此，cdc2诱导的vimentin-Ser55磷酸化导致了这种升高。通过RNA干扰减少Plkl可破坏有丝分裂中vimentin-Ser82磷酸化的升高。这些结果表明，Cdc2不仅通过直接的酶反应，还通过Plkl募集到vimentin，从而调控有丝分裂期vimentin磷酸化的新机制。2)我们发现Cdc2磷酸化内着丝粒蛋白（inner centromere protein, cenp）上的Thr59和Thr388，从前期到中期调控Aurora-B的定位和激酶活性。enimp的耗尽会破坏Plkl在着丝点的定位。这种表型由外源表达的INCENP野生型（WT）拯救，并且在Thr59到Ala （T59A）突变，而在Thr388到Ala （T388A）突变。用T388A替代内源性的INCENP导致从中期到后期的进展延迟。我们的研究结果表明，Cdc2对INCENP的磷酸化是Plkl向着丝点募集的必要条件，Plkl和Aurora-B在INCENP上的复合物形成可能在染色体动力学调节中起关键作用。

项目成果

期刊论文数量（113）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Complex formation of Plkl and INCENP required for metaphase- anaphase transition.

中期-后期转变所需的 Plkl 和 INCENP 的复杂形成。

DOI：
发表时间：
2006
期刊：
Nature Cell Biology 8
影响因子：
0
作者：
Goto;H. et al.
通讯作者：
H. et al.

Kawajiri, A., et al.: "Functional significance of the specific sites phosphorylated in desmin at cleavage furrow : Aurora-B may phosphorylate and regulate type III intermediate filaments during cytokinesis coordinatedly with Rho-kinase."Mol.Biol.Cell. 14.

Kawajiri, A. 等人：“卵裂沟结蛋白中特定位点磷酸化的功能意义：Aurora-B 可能在胞质分裂过程中与 Rho 激酶协调磷酸化和调节 III 型中间丝。”Mol.Biol.Cell。

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

Regulation of mitotic function of Chk1 through phosphorylation at novel sites by cyclin-dependent kinase 1 (Cdk1)

DOI：
10.1111/j.1365-2443.2006.00955.x
发表时间：
2006-05-01
期刊：
GENES TO CELLS
影响因子：
2.1
作者：
Shiromizu, T;Goto, H;Inagaki, M
通讯作者：
Inagaki, M

PKCepsilon-mediated phosphorylation of vimentin controls integrin recycling and motility.

DOI：
发表时间：
2005
期刊：
The EMBO journal
影响因子：
0
作者：
J. Ivaska;Karoliina Vuoriluoto;T. Huovinen;I. Izawa;M. Inagaki;P. Parker
通讯作者：
J. Ivaska;Karoliina Vuoriluoto;T. Huovinen;I. Izawa;M. Inagaki;P. Parker

Vimentin‐Ser82 as a memory phosphorylation site in astrocytes

DOI：
10.1111/j.1365-2443.2006.00961.x
发表时间：
2006-05
期刊：
Genes to Cells
影响因子：
2.1
作者：
Takashi Oguri;Akihito Inoko;H. Shima;I. Izawa;N. Arimura;Tomoya Yamaguchi;N. Inagaki;K. Kaibuchi;K. Kikuchi;M. Inagaki
通讯作者：
Takashi Oguri;Akihito Inoko;H. Shima;I. Izawa;N. Arimura;Tomoya Yamaguchi;N. Inagaki;K. Kaibuchi;K. Kikuchi;M. Inagaki