RIP1磷酸化调控细胞死亡信号转导复合体Ripoptosome形成机制的研究

项目名称： RIP1磷酸化调控细胞死亡信号转导复合体Ripoptosome形成机制的研究

项目编号： No.31201043

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 遗传学与生物信息学、细胞生物学

项目作者： 葛源

作者单位： 中国海洋大学

项目金额： 23万元

中文摘要： Ripoptosome是在哺乳动物肿瘤细胞中新发现的一个细胞死亡信号复合体。它的形成和下游caspase激活依赖于核心组分RIP1的磷酸激酶活性。据报道，RIP1磷酸激酶活性受RIP1磷酸化调控。但RIP1磷酸化、RIP1磷酸激酶活性与Ripoptosome形成之间的关系并不明确，这也是本课题的核心内容。本课题拟使用分子筛、免疫共沉淀、定点突变、RNAi、磷酸化免疫印迹等手段，研究RIP1磷酸化对RIP1磷酸激酶活性、Ripoptosome形成和细胞死亡的作用，分析RIP1磷酸化对自身稳定性的影响，探究分子伴侣Hsp90保护磷酸化RIP1的机制以及RIP1磷酸化与FADD磷酸化的关系。本项申请有望在RIP1调控Ripoptosome形成、RIP1与FADD相互识别、Ripoptosome稳定性调控机制等方面取得创新性成果，为更好地阐述细胞死亡通路奠定基础，为肿瘤病理和药物研发提供依据。

中文关键词： 受体相互作用蛋白丝氨酸/苏氨酸激酶1；细胞死亡；磷酸化；促细胞死亡蛋白质复合体Ripoptosome；

英文摘要： Ripoptosome is a newly discovered mammalian intracellular signalling complex involved in cell death pathways. Its formation and subsequent caspase activation depends on RIP1 kinase activity. As reported previously, RIP1 kinase activity is regulated by its phosphorylation. However, the machinery among RIP1 phosphorylation, kinase activity and Ripoptosome formation is still unknown. It therefore becomes the main component in this proposal. In this proposal, relevant molecular and cellular methods including gel filtration, immunoprecipitation, point mutagenesis, RNAi, western blot for phophorylated protein are outlined to further investigate the effect of RIP1 phosphorylation on its kinase activity, Ripoptosome formation, cell fate as well as RIP1 self stability. Role of the molecular chaperon, Hsp90 in protecting phosphorylated RIP1 from degradation and relationship between RIP1 phosphorylation and FADD phosphorylation will also be studied. Innovative discovery is hopefully to be obtained in the mechanism of Ripoptosome formation regulated by RIP1, RIP1 and FADD recognition to initiate Ripoptosome and Ripoptosome half-life control. This research will eventually further our understanding in cell death signalling pathway, tumour pathological knowledge and benefit the innovation of anti-cancer drugs.

英文关键词： RIP1；cell death；phosphorylation；pro-cell death complex Ripoptosome；