死亡相关蛋白激酶蛋白降解调控通路的研究

项目名称： 死亡相关蛋白激酶蛋白降解调控通路的研究

项目编号： No.31301172

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 生物科学

项目作者： 林尧

作者单位： 福建师范大学

项目金额： 23万元

中文摘要： 死亡相关蛋白激酶 (DAPK-1) 是一种丝氨酸-苏氨酸激酶，在细胞凋亡、自噬、增殖等生理过程中具有重要作用。DAPK-1蛋白同时受到了蛋白酶体与溶酶体降解通路的双重调控，但目前对其调控机制的研究仅限于蛋白酶体通路中的E3上。本项目将利用泛素或类泛素小分子E1和E2的siRNA库，探寻影响DAPK-1蛋白降解的新信号通路及其作用机制，同时也将研究蛋白酶体与溶酶体两条蛋白降解通路之间的互动（crosstalk）。此外DAPK-1基因的甲基化及随之而来的mRNA表达缺失在许多癌症中都有报导，但对与之相关的蛋白表达却所知甚少。我们将在慢性淋巴白血病的原发性癌症样本中考查DAPK-1的蛋白表达，并与其mRNA表达进行对比，探讨蛋白降解通路在癌症中对抗癌基因表达调控的重要性。我们的研究将有助于更全面地理解DAPK-1蛋白的降解，完善其分子调控网络，并为DAPK-1在癌症中的表达调控提供更深入的理解。

中文关键词： 死亡相关蛋白激酶；蛋白降解；类泛素；溶酶体；DNA 甲基化

英文摘要： Death Associated Protein Kinase 1 (DAPK-1) is a serine/threonine kinase involved in the regulation of multiple cellular events such as apoptosis, autophagy and proliferation. Protein degradation is critical for controlling DAPK-1level and it has been found that DAPK-1 is degraded through both ubiquitin-proteasomal and lysosomal degradation pathways. In this project, we aim to search new regulatory signalling pathways controlling DAPK-1 protein degradation by using a siRNA library of E1 and E2 enzymes for ubiquitin and ubiquitin-like molecules. Once the relevant E1s or E2s are identified, we plan to investigate the mechanisms underlying their regulation of DAPK-1 degradation, which may also provide new insight into the crosstalk between proteasome and lysosome. In addition, DNA methylations of DAPK-1 gene and consequent mRNA loss have been discovered in various tumours. However, it is not clear whether DAPK-1 protein expression is also lost in these tumours. We plan to investigate the DAPK-1 protein levels in primary tumour samples that are known to be low or null in DAPK-1 mRNA expression. If DAPK-1 protein is detected in these samples, we will investigate the mechanisms for the imbalance of mRNA and protein, which will start from the degradation pathways. Our results will provide a broader understanding of DAPK

英文关键词： dapk-1；Protein degradation；Ubiquitin-like；Lysosome；DNA methylation