Role of MAP kinase cascades in the regulation of diverse cellular functions

MAP 激酶级联在调节多种细胞功能中的作用

基本信息

批准号：
17390020
负责人：
KOHNO Michiaki
金额：
$ 9.6万
依托单位：
Nagasaki University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17390020/
关键词：
ERK-MAP Kinase Intracellular Localization GEF-H1 Cell Motility c-Jun N-terminal Kinase Intermediate Filaments Cell Cycle Spindle Checkpoint スピンドルチェックポイント p90^<RSK>RhoA 細胞内局在性ケラチン8 細胞質分裂

项目摘要

1. We have examined a possible molecular mechanism through which nuclear translocation and retention of ERK-MAP kinases is regulated. A novel 26-kD protein (p26) which contains a SH3 domain at the N-terminus and three ankyrin repeat sequences at the C-terminus has been identified as a candidate molecule which is involved in the regulation of nuclear translocation/retention of ERK-MAP kinases. Overexpression of p26 suppresses the HGF-induced nuclear localization/retention of ERK-MAP kinases in MDCK cells, while siRNA-mediated knockdown of p26 enhances it. p26 interact specifically with a novel 120-kDa protein (p120), and this interaction is suppressed by the phosphorylation of p26 by p90^<rsk>, an effector molecule downstream of the ERK-MAP kinases. These results suggest that nuclear translocation/retention of ERK-MAP kinases is regulated by the ERK-MAP kinase signaling pathway by itself.2. A novel GDP/GTP exchanger of RhoA, GEF-H1, has been shown to be a physiological substrate of ERK- … More MAP kinases. Expression of GEF-H1 is up-regulated by the ERK MAP kinase pathway. Phosphorylation of Thr^<678> by ERK-MAP kinases induces the activation of GEF-H1 thereby activates RhoA, whereas it induces the inhibition of Rac1. Furthermore, siRNA-mediated knock down of GEF-H1 enhances the cell motility response. These results suggest that GEF-H1 is involved in the ERK-MAF kinase pathway-mediated cell motility response.3. c-Jun N-Terminal Kinase (JNK) has been shown to be involved in the regulation of cytokinesis. JNK phosphorylates keratin 8 to induce the relaxation of keratin filaments, which appear to be one of the prerequisites for cells to undergo cytokinesis.4. Combination of 50 μM PD98059 and a low concentration (3 nM) of vincristin induces marked apoptotic cell death response in G_2/M-phase-arrested T24 cells, but not in G_1-/S-phase-arrested cells. Under such conditions, accumulation of cyclinB, Plk1and Aurora-B has been observed. These results suggest that ERK-MAP Kinase pathway is involved in the regulation of spindle check point. Less

1. 我们研究了一种可能的分子机制，通过该机制调节 ERK-MAP 激酶的核转位和保留。一种新型 26-kD 蛋白 (p26)，其 N 末端包含一个 SH3 结构域，C 末端包含三个锚蛋白重复序列。 -末端已被确定为参与 ERK-MAP 激酶核易位/保留调节的候选分子，p26 的过度表达可抑制 HGF 诱导的核。 ERK-MAP 激酶在 MDCK 细胞中的定位/保留，而 siRNA 介导的 p26 敲除增强了 p26 与新型 120-kDa 蛋白 (p120) 的特异性相互作用，并且这种相互作用被 p90^< 的 p26 磷酸化所抑制。 rsk>，ERK-MAP 激酶下游的效应分子。这些结果表明 ERK-MAP 激酶的核转位/保留是存在的。 2. RhoA 的一种新型 GDP/GTP 交换剂 GEF-H1 已被证明是 ERK-... 更多 MAP 激酶的生理底物。 -由ERK MAP激酶途径调节Thr^<678>通过ERK-MAP激酶的磷酸化诱导GEF-H1的激活，从而激活RhoA，而它此外，siRNA 介导的 GEF-H1 敲低可增强细胞运动反应。这些结果表明，GEF-H1 参与了 ERK-MAF 激酶途径介导的细胞运动反应。3． N 末端激酶 (JNK) 已被证明参与胞质分裂的调节，JNK 磷酸化角蛋白 8，从而诱导角蛋白丝松弛。似乎是细胞进行胞质分裂的先决条件之一。4. 50 μM PD98059 和低浓度 (3 nM) 长春新碱的组合可在 G_2/M 期停滞的 T24 细胞中诱导明显的凋亡细胞死亡反应。在这样的条件下，G_1-/S-期停滞的细胞，观察到了cyclinB、Plk1和Aurora-B的积累。表明ERK-MAP激酶通路参与纺锤体检查点的调节较少。