Analysis of a novel gene DP5 that induces programmed cell death

诱导程序性细胞死亡的新基因 DP5 的分析

• 批准号: 10480219
• 负责人: WANAKA Akio
• 金额: $ 8.9万
• 依托单位: Fukushima Medical University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10480219/
• 关键词: Apoptosis; DP5; Bcl-2; BH3 domain; neuron; amyotrophic lateral sclerosis; 神経細胞; 強制発現; Yeast Two hybrid; Bcl-2 ファミリー; BH3 ドメイン; 脳虚血

We cloned a novel gene, whose expression was upregulated when cells undergo apoptosis, and named DP5. DP5 gene encoded a protein consisting of 92 amtino acids. DP5 protein had a BH3 (Bcl-2 homology domain 3) domain and had apoptosis-inducing activity when overexpressed in cultured neurons. These results suggested that DP5 was a novel proapoptotic member of the Bcl-2 family. We investigated distribution pattern of DP5 mRNA in the developing rat embryo by in situ hybridization and found that it was specifically expressed in the embryonic spinal motoneurons and autonomic ganglia. These tissues are known to prune superfluous neurons by apoptotic mechanism. Therefore, the DP5 gene might be involved in such apoptotic process. We also detected moderate DP5 mRNA expression in the postnatal hippocampus and cerebral cortex. We noted significant upregulation of the DP5 gene expression in the adult hippocampus aud cerebral cortex in response to ischemic insults, suggesting that DP5 is also involved in the ischemic neuronal death. Recent studies revealed an increasing number of BH3 only proteins such as Bik, Bim, and Blk. Among them, Bim specifically interacted with cytoskeletal component. This interaction regulated Bim's cell-death inducing activity ; detachment from cytoskeleton promoted Bim to relocate to mitochondrial membrane, and thereby induced apoptosis. Such a regulatory mechanisms may exist in the case of DP5. We searched for DP5-interacting molecule with yeast-two hybrid method. We have so far found three candidates, one of which is hn RNP.Possible regulatory mechanisms by these candidates are to be investigated in near future. Finally we investigated possible involvement of DP5 in degenerative neurological disease. We found that Harakiri (human homolog of DP5) was specifically expressed in the motoneurons of the amyotrophic lateral sclerosis patients.

我们克隆了一个在细胞凋亡时表达上调的新基因，命名为DP5。DP5基因编码了一种由92个氨基酸组成的蛋白质。DP5蛋白具有BH3 （Bcl-2同源结构域3）结构域，在培养的神经元中过表达时具有诱导凋亡活性。这些结果表明DP5是Bcl-2家族中一个新的促凋亡成员。我们通过原位杂交研究DP5 mRNA在发育中的大鼠胚胎中的分布规律，发现它在胚胎脊髓运动神经元和自主神经节中特异性表达。已知这些组织通过凋亡机制修剪多余的神经元。因此，DP5基因可能参与了这一凋亡过程。我们还检测到DP5 mRNA在出生后海马和大脑皮层中有中度表达。我们注意到成年海马和大脑皮层中DP5基因表达在缺血损伤后显著上调，这表明DP5也参与了缺血性神经元死亡。最近的研究表明，越来越多的BH3蛋白，如Bik、Bim和Blk。其中，Bim与细胞骨架组分特异性相互作用。这种相互作用调节了Bim的细胞死亡诱导活性；脱离细胞骨架促使Bim迁移至线粒体膜，从而诱导细胞凋亡。在DP5的情况下可能存在这样的监管机制。用酵母- 2杂交法寻找dp5相互作用分子。到目前为止，我们已经找到了三个候选人，其中一个是RNP。这些候选国家可能的监管机制将在不久的将来进行调查。最后，我们调查了DP5在退行性神经疾病中的可能参与。我们发现Harakiri （DP5的人类同源物）在肌萎缩性侧索硬化症患者的运动神经元中特异性表达。

项目成果

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