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Molecular mechanisms of response of living body to environmental stress

生命体对环境应激反应的分子机制

基本信息

批准号：
14370050
负责人：
TAMURA Shinri
金额：
$ 7.55万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370050/
关键词：
stress SAPK phosphatase gene knockout mouse ES cell BMPR II scaffold protein

项目摘要

1.We investigated the role(s) of PP2Cε in the regulation of IL-1 signaling pathways. Ectopic expression of PP2Cε inhibited the IL-1-and TAK1-induced activation of mitogen-activated protein kinase kinase 4 (MKK4)-c-Jun N-terminal kinase or MKK3-p38 signaling pathway. PP2Cε dephosphorylated TAK1 in vitro. Co-immunoprecipitation experiments indicated that PP2Cε associates stably with TAK1 and attenuates the binding of TAK1 to MKK4 or MKK6. Ectopic expression of a phosphatase-negative mutant of PP2Cε, PP2Cε (D/A), which acted as a dominant negative form, enhanced both the association between TAK1 and MKK4 or MKK6 and the TAK1-induced activation of an AP-1 reporter gene. The association between PP2Cε and TAK1 was transiently suppressed by IL-1 treatment of the cells. Taken together, these results suggest that, in the absence of IL-1-induced signal, PP2Cε contributes to keeping the TAK1 signaling pathway in an inactive state by associating with and dephosphorylating TAK1.2.We investigated th … More e physiologic functions of PP2Cβ using a gene knockout technique in mice. We found that most PP2Cβ^<Δ/Δ> embryos die between the two-and eight-cell stages, whereas PP2Cβ^<Δ/Δ> ES cells are viable, suggesting that relatively high PP2Cβ expression is required for the early stages of pre-implantation development. PP2Cβ knockdown studies using ES cells revealed that serum-induced activation of p38, but not JNK, ERK, or Akt, was enhanced further by decreased PP2Cβ expression. Since p38 has been implicated in the negative regulation of mitosis in early pre-implantation embryos, these observations raise the possibility that cell division at early cleavage stages might be severely disturbed due to the enhanced activity of p38 in PP2Cβ^<Δ/Δ> embryos.3.We have shown that BMPRII, a BMP type II receptor, associates with JNK through its unique carboxyl-terminal region, which contains a putative JNK binding domain. BMPRII was also able to bind to MKK4, MKK7 and ASK1. BMP2 treatment of cells induced the recruitment of JNK to BMPRII and activated the ASK1 signaling module associated with BMPRII. These results suggest that BMPRII participates in the spatial regulation of the ASK1-JNK signaling pathway as a cell-membrane associated scaffolding protein. Less

1.我们研究了PP2Cε在调节IL-1信号通路中的作用。PP2Cε的异位表达抑制il -1和tak1诱导的丝裂原活化蛋白激酶激酶4 (MKK4)-c-Jun n-末端激酶或MKK3-p38信号通路的激活。PP2Cε在体外使TAK1去磷酸化。共免疫沉淀实验表明，PP2Cε与TAK1稳定结合，并减弱TAK1与MKK4或MKK6的结合。PP2Cε磷酸酶阴性突变体PP2Cε (D/ a)的异位表达作为显性阴性形式，增强了TAK1与MKK4或MKK6之间的关联以及TAK1诱导的AP-1报告基因的激活。细胞IL-1处理可暂时抑制PP2Cε和TAK1之间的关联。综上所述，这些结果表明，在缺乏il -1诱导信号的情况下，PP2Cε通过与TAK1.2相关联并使其去磷酸化，有助于保持TAK1信号通路处于非活性状态。我们利用基因敲除技术研究了PP2Cβ在小鼠体内的生理功能。我们发现大多数PP2Cβ^<Δ/Δ>胚胎在2 - 8细胞阶段死亡，而PP2Cβ^<Δ/Δ> ES细胞是有活力的，这表明相对较高的PP2Cβ表达是植入前早期发育所必需的。利用ES细胞进行的PP2Cβ敲除研究表明，血清诱导的p38激活，而不是JNK， ERK或Akt，通过降低PP2Cβ表达进一步增强。由于p38与早期着床前胚胎有丝分裂的负调控有关，这些观察结果提出了一种可能性，即由于p38在PP2Cβ^<Δ/Δ>胚胎中的活性增强，早期卵裂阶段的细胞分裂可能受到严重干扰。我们已经证明BMP II型受体BMPRII通过其独特的羧基末端区域与JNK结合，该区域包含假定的JNK结合域。BMPRII也能与MKK4、MKK7和ASK1结合。BMP2处理细胞诱导JNK向BMPRII募集，并激活与BMPRII相关的ASK1信号传导模块。这些结果表明，BMPRII作为细胞膜相关支架蛋白参与ASK1-JNK信号通路的空间调控。少