Identification and functional analysis of Cdk5-mediated phosphorylation of the proteins that are related to brain formation and development

Cdk5介导的与大脑形成和发育相关的蛋白质磷酸化的鉴定和功能分析

• 批准号: 15500273
• 负责人: OHSHIMA Toshio
• 金额: $ 2.37万
• 依托单位: RIKEN
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15500273/
• 关键词: phosphorylation; serine; threonine kinase; cyclin-dependent kinase; GSK3β; tubulin; Reelin; Dab1; Alzheimer's disease; 大脳皮質; 細胞移動

Purpose and Method :Cdk5 is a neuron-specific Ser/Thr protein kinase and Cdk5 KO mice exhibit defective brain development particularly in layer formation of cortical structures. Abnormalities of brain development in Cdk5 KO mice are caused by defects in the phosphorylations of brain development related-proteins. Using brains from Cdk5 KO mice, we conducted a screening to identify Cdk5 substrate(s). Functional analysis of newly identified substrate(s) is further performed. We also studied the functional significance of Cdk5-mediated phosphorylation of Dab1 which is a intracellular mediator of Reelin signaling.Results :Through the screening, we identified CRMP2 as a substrate of Cdk5. Under the collaboration with Dr.Goshima's group in Yokohama City University School of Medicine, we found Cdk5 phosphorylates CRMP2 at Ser522. This phosphorylation is required for Ser509 phosphorylation by GSK3β. This sequential phosphorylation by Cdk5 and GSK3β reduces the affinity between CRMP2 and tubulins. We also demonstrate that this sequential phosphorylation of CRMP2 is related to the production of 3F4 reactivity specifically in the brains from Alzheimer's patients. In another study, we showed that Cdk5 phosphorylates Dab1 at multiple sites in its C-terminal. Cdk5-mediated Ser/Thr phosphorylations of Dab1 inhibit tyrosine phosphorylation of Dab1 induced by Reelin. This result indicates a negative modulation of Reelin signaling by Cdk5.

目的和方法：Cdk5 是一种神经元特异性 Ser/Thr 蛋白激酶，Cdk5 KO 小鼠表现出大脑发育缺陷，特别是在皮质结构的层形成方面。 Cdk5 KO小鼠大脑发育异常是由大脑发育相关蛋白磷酸化缺陷引起的。使用 Cdk5 KO 小鼠的大脑，我们进行了筛选以鉴定 Cdk5 底物。进一步对新鉴定的底物进行功能分析。我们还研究了Cdk5介导的Dab1磷酸化的功能意义，Dab1是Reelin信号传导的细胞内介质。结果：通过筛选，我们鉴定了CRMP2作为Cdk5的底物。在与横滨市立大学医学院Goshima博士团队的合作下，我们发现Cdk5在Ser522处磷酸化CRMP2。 GSK3β 磷酸化 Ser509 需要这种磷酸化。 Cdk5 和 GSK3β 的这种连续磷酸化降低了 CRMP2 和微管蛋白之间的亲和力。我们还证明，CRMP2 的这种顺序磷酸化与 3F4 反应性的产生有关，特别是在阿尔茨海默病患者的大脑中。在另一项研究中，我们发现 Cdk5 在其 C 端的多个位点磷酸化 Dab1。 Cdk5 介导的 Dab1 Ser/Thr 磷酸化抑制 Reelin 诱导的 Dab1 酪氨酸磷酸化。该结果表明 Cdk5 对 Reelin 信号传导进行负调节。

项目成果

Modulation of Reelin signaling by cyclin-dependent kinase 5

• DOI: 10.1016/j.brainres.2006.01.121
• 发表时间: 2007-04-06
• 期刊: BRAIN RESEARCH
• 影响因子: 2.9
• 作者: Ohshima, Toshio;Suzuki, Hiromi;Mikoshiba, Katsuhiko
• 通讯作者: Mikoshiba, Katsuhiko

Semaphorin-3A signaling is mediated via sequential Cdk5-GSK3B phosphorylation of CRMP2 : Impli-sation of common phoshorylating mechanism underlying axon guidance and Alzheimer's disease

Semaphorin-3A 信号传导通过 CRMP2 的连续 Cdk5-GSK3B 磷酸化介导：轴突引导和阿尔茨海默氏病的常见磷酸化机制的暗示

• 发表时间: 2005
• 期刊: Genes to Cells Vol.10,Issue 2
• 作者: Takakusaki K.;et al.;Y.Uchida ら
• 通讯作者: Y.Uchida ら

Cdk5-dependent regulation of glucose-stimulated insulin secretion

• DOI: 10.1038/nm1299
• 发表时间: 2005-10-01
• 期刊: NATURE MEDICINE
• 影响因子: 82.9
• 作者: Wei, FY;Nagashima, K;Tomizawa, K
• 通讯作者: Tomizawa, K

Semaphorin-3A signaling is mediated via sequential Cdk5-GSK3β phosphorylation of CRMP2 : implication of common phosphorylating mechanism underlying axon guidance and Alzheimer's disease.

Semaphorin-3A 信号传导通过 CRMP2 的连续 Cdk5-GSK3β 磷酸化介导：轴突引导和阿尔茨海默病的常见磷酸化机制的暗示。

• 发表时间: 2005
• 期刊: Genes to Cells 10
• 作者: Uchida;Y.;Ohshima;T.;Sasaki;Y.;Suzuki;H.;Yanai;S.;Yamashita;N.;Nakamura;F.;Takei;K.;Ihara;Y.;Mikoshiba;K.;Kolattukudy;P.;Honnorat;J.;Goshima;Y.
• 通讯作者: Y.

M.Hirasawa, T.Ohshima, S.Takahashi et al.: "Perinatal abrogation of Cdk5 expression in brain results in neuronal migration defects"Proceeding National Academy of Science U.S.A.. 印刷中. (2004)

M.Hirasawa、T.Ohshima、S.Takahashi 等人：“脑中 Cdk5 表达的围产期废除导致神经元迁移缺陷”美国国家科学院院刊，出版中（2004 年）。