Identification and characterization of CF kinase

CF 激酶的鉴定和表征

• 批准号: 08458234
• 负责人: INAGAKI Masaki
• 金额: $ 3.84万
• 依托单位: Aichi Cancer Center
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08458234/
• 关键词: Intermediate filament; Cytokinesis; Rho-kinase; CF-kinase; Phosphorylation; 細胞周期; 細胞質分裂; 中間径フィラメント; キナーゼ; リン酸化ペプチド抗体; 低分子量GTP結合蛋白質; 蛋白質リン酸化反応

Cytokinesis is the mitotic precess in which all the essential cell components are segregated into postmitotic daughter cells It is suggested that protein phoshorylation plays pivotal roles in mitotic control, however the mechanisms for the cellular separation in cytokineses are not well characterized. We reported a protein kinase activity that phosphorylates a type III intermediate filament protein, glial fiberllary acidic protein (GFAP), specifically at the cleavage furrow of cytokinetic cells. It was termed cleavage furrow (CF) kinase. Now, we obtained evidence that Rho-associated Kinase (Rho-kinase) is a candidate for CF kinase ; Rho-kinase phosphorylated GFAP in vitro at the same sites that were phosphorylated by CF kinase in vivo. We also analyzed the functions of CF kinase/Rho-kinase activity by introducing mutant GFAPs into type III intermediate filament-negative cells. The mutation in CF kinase/Rho-kinase phosphorylation sites impaired GFAP segregation into postmitotic daughter cells. As a result, unusually long cytoplasmic bridge-like structure was formed between unseparated dughter cells. These results suggst that CF kinase/Rho-kinase is essential for the segragation of GFAP into daughter cells which in turn ensures efficient cellular separation.

细胞因子是有丝分裂的动物，其中所有必需的细胞成分都被隔离为有丝分裂后子细胞，建议蛋白质磷酸化在有丝分裂控制中起关键作用，但是细胞因子中细胞分离的机制并未很好地表征。我们报道了一种蛋白激酶活性，该蛋白激酶活性磷酸化III型中间丝蛋白，神经胶质酸性蛋白（GFAP），特别是在细胞动物细胞的裂解沟中。它被称为裂解沟（CF）激酶。现在，我们获得了与Rho相关激酶（Rho-激酶）是CF激酶的候选者的证据。在体内CF激酶磷酸化的相同位点，Rho-kinase磷酸化的GFAP体外。我们还通过将突变型GFAP引入III型中间丝 - 阴性细胞中，分析了CF激酶/Rho-激酶活性的功能。 CF激酶/Rho-激酶磷酸化位点中的突变损害了GFAP分离到有丝分裂后子细胞中。结果，在未分离的dughter细胞之间形成了异常长的细胞质桥样结构。这些结果表明，CF激酶/Rho-激酶对于将GFAP脱离为子细胞至关重要，这又确保了有效的细胞分离。

项目成果

Inagaki, N., Goto, H., Ogawara, M., Nishi, Y., Ando, S., and Inagaki, M.: "Spatial patterns of Ca^<2+> signals define the intracellular distribution of a signaling by Ca^<2+>/calmodulin-dependent protein kinase II." J.Biol.Chem.272. 25195-25199 (1997)

Inagaki, N.、Goto, H.、Okawara, M.、Nishi, Y.、Ando, S. 和 Inagaki, M.：“Ca^<2> 信号的空间模式定义了 Ca 信号传导的细胞内分布。

Sekimata,M: "Detection of protein kinase activity specifically activated at metaphase-anaphase transition." J.Cell Biol.132. 635-641 (1996)

Sekimata，M：“检测在中期-后期转变时特异激活的蛋白激酶活性。”

Hosako, H., Amano, M., and Inagaki, M.et al.: "Phosphorylation of glial fibrillary acidic protein at the same sites by cleavage furrow kinase and Rho-associated kinase." J.Biol.Chem. 272. 10333-10336 (1997)

Hosako, H.、Amano, M. 和 Inagaki, M. 等人：“通过裂解沟激酶和 Rho 相关激酶在相同位点磷酸化神经胶质原纤维酸性蛋白。”

Inagaki,M.: "Dynamic property of intermediate filaments: Regulation by phosphorylation." BioEssays. 18. 481-487 (1996)

Inagaki,M.：“中间丝的动态特性：磷酸化调节。”

Inagaki,M.,Matsuoka,Y.,and Inagaki,N.et al.: "Dynamic property of intermediate filaments : Regulation of phosphorylation." BioEssays. 18. 481-487 (1996)

Inagaki，M.，Matsuoka，Y.，and Inagaki，N.等人：“中间丝的动态特性：磷酸化的调节。”