权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Identification and characterization of scaffold porteis in JNK cascades

JNK 级联中门脉支架的鉴定和表征

基本信息

批准号：
13680777
负责人：
YOSHIOKA Katsuji
金额：
$ 2.24万
依托单位：
Kanazawa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13680777/
关键词：
MAP kinase Signal transduction mouse embryonic stem cell Scaffold protein 細胞内シグナル伝達 JNK

项目摘要

Scaffold proteins in MAP kinase(MAPK) signal transduction pathways organize MAPK signaling components into functional MAPK modules, thereby enabling the efficient and specific activation of MAPK pathways. We previously identified two novel JNK MAPK-binding proteins, termed JSAP1 and JNKBP1, which may function as scaffolding proteins in the MAPK pathways. In this research project, we have further analyzed the putative scaffold proteins, and obtained the following results. 1.We have identified family members of JSAP1 and JNKBP1,termed JSAP2 and JNKBP2,respectively. 2.We have analyzed the functional relationship between JSAP1 and ASK MAPKKK that activates JNK and p38 MAPK pathways in response to environmental stresses such as reactive oxygen species. Our data suggest that JSAP1 dynamically participates in signal transduction by forming a phosphorylation-dependent signaling complex in the ASK-JNK signaling module. 3.We established JSAP1-null mouse embryonic stem(ES) cell lines. Neurite outgrowth from the JSAP1-null embryoid bodies was apparently less efficient than from wild type. We also observed altered expression of differentiation markers, especially markers for neuroectoderm during in vitro differentiation of JSAP1-null mutant. These results suggest that JSAP1 may be required for early embryonic neurogenesis. 4.We established stable PC12h cell lines that expressed wild-type JSAP1 and its mutant lacking the JNK-binding domain(JBD). immunocytochemical studies of the cell lines indicated that the mutant JSAP1 was localized to the growth cones of differentiating PC12h cells in a similar manner to wild-type JSAP, suggesting that the proper subcellular localization of JSAP1 along microtubules probably does not require INK signaling.

MAP激酶（MAPK）信号转导通路中的支架蛋白将MAPK信号组分组织成功能性MAPK模块，从而使得MAPK通路能够有效和特异性地活化。我们以前确定了两个新的JNK MAPK结合蛋白，称为JSAP 1和JNKBP 1，这可能是MAPK通路中的支架蛋白。在本研究项目中，我们进一步分析了推定的支架蛋白，并获得了以下结果。1.我们鉴定了JSAP 1和JNKBP 1的家族成员，分别命名为JSAP 2和JNKBP 2。2.我们分析了JSAP 1和ASK MAPKKK之间的功能关系，ASK MAPKKK激活JNK和p38 MAPK通路，响应环境应激，如活性氧。我们的数据表明，JSAP 1动态参与信号转导，形成磷酸化依赖的信号复合物中的ASK-JNK信号模块。3.建立了JSAP 1基因敲除的小鼠胚胎干细胞系。JSAP 1缺失胚状体的神经突生长效率显然低于野生型。我们还观察到改变表达的分化标志物，特别是标记的神经外胚层在体外分化的JSAP 1无效突变体。这些结果表明，JSAP 1可能是早期胚胎神经发生所必需的。4.建立了稳定表达野生型JSAP 1及其缺失JNK结合域突变体（JBD）的PC 12 h细胞系。细胞系的免疫细胞化学研究表明，突变型JSAP 1以与野生型JSAP相似的方式定位于分化中的PC 12 h细胞的生长锥，这表明JSAP 1沿沿着微管的适当亚细胞定位可能不需要INK信号传导。