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Proteome analysis of phosphorylation signaling pathways involved in maxillofacial development

颌面发育相关磷酸化信号通路的蛋白质组分析

基本信息

批准号：
15390559
负责人：
TAKEDA Kohsuke
金额：
$ 8.26万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15390559/
关键词：
Phosphorylation Signal transduction Proteome Morphogenesis

项目摘要

p38 MAP kinase pathway is an intracellular signal transduction pathway using a phosphorylation relay and is involved in the regulation of apoptosis during development and differentiation. We have established a cell culture system, in which neuronal differentiation of PC12 cells is induced by the selective and constitutive activation of p38 MAP kinase through the expression of constitutively active ASK1, an activator of the p38 pathway. Here we have identified oncoprotein 18 (Op18)/stathmin as a novel potential target of the p38 cascade. By two-dimensional electrophoresis, phosphorylation of Op18/stathmin was found to be increased upon the expression of constitutively active ASK1 in PC12 cells. The ASK1-dependent increase in the phosphorylation of Op18/stathmin was attenuated by the treatment with SB203580, suggesting that p38α and/or p38β contribute to the phosphorylation of Op18/stathmin. Consistently, we found that all four isoforms of p38 directly phosphorylated Op18/stathmin primarily at serine 25 in vitro. Taken together with the quantitative RT-PCR data indicating that p38α was the dominantly expressed isoform in PC12 cells, ASK1-induced phosphorylation of Op18/stathmin appears to be mediated mainly through p38α in these cells. Given that the microtubule-destabilizing activity of Op18/stathmin is regulated by its phosphorylation, the ASK1-p38 cascade may regulate microtubule dynamics through Op18/stathmin.

p38 MAP激酶通路是一种利用磷酸化中继的细胞内信号转导通路，参与发育和分化过程中细胞凋亡的调控。我们已经建立了一个细胞培养系统，在该系统中，PC12细胞的神经元分化是由p38 MAP激酶的选择性和组成性激活（通过表达组成性活性ASK1， p38通路的激活因子）诱导的。在这里，我们已经确定了癌蛋白18 (Op18)/stathmin作为p38级联的一个新的潜在靶点。通过双向电泳，发现PC12细胞中组成活性ASK1的表达增加了Op18/stathmin的磷酸化。用SB203580处理后，ask1依赖性的Op18/stathmin磷酸化增加被减弱，这表明p38α和/或p38β参与了Op18/stathmin的磷酸化。一致地，我们发现p38的所有四种亚型在体外直接磷酸化Op18/stathmin，主要是在丝氨酸25处。结合定量RT-PCR数据显示p38α是PC12细胞中主要表达的亚型，ask1诱导的Op18/stathmin磷酸化似乎主要通过p38α介导。鉴于Op18/stathmin的微管不稳定活性受其磷酸化调控，ASK1-p38级联可能通过Op18/stathmin调节微管动力学。