A genetic analysis on a novel regulatory mechanism to induce cell death in response to perturbation of dynamic instability of microtubules

对微管动态不稳定性扰动诱导细胞死亡的新型调控机制的遗传分析

• 批准号: 17570006
• 负责人: INOUE Yoshihiro
• 金额: $ 2.3万
• 依托单位: Kyoto Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17570006/
• 关键词: microtubules; Drosophila; apoptosis; Orbit; dynamic instability

Microtubules are indeed involved in many cellular phenomena. It has been shown that dynamic instability of microtubules is especially important for regulation of some cellular events. The Orbit/CLASP, a conserved microtubule-associated protein enriched at plus-ends of microtubules plays an essential role in regulation of the dynamic instability. Loss-of-function mutations of the orbit in Drosophila increase instability of microtubules. Conversely, over-expression of the gene in imaginal discs using GAL4-UAS system causes excessive stabilization of microtubules and consequently a formation of microtubule bundles that are never observed in normal cells at interphase. The over-expression of the orbit in eye disc cells after morphogenic furrows accumulates metaphase cells, suggesting it activates the microtubule assembly checkpoint in mitosis. We found that the over-expression eventually induces apoptosis dependent on Caspase 3 at post-mitotic stage in the eye discs. Knockdown of the gene also causes induction of the Caspase 3 in wing discs. This type of apoptosis cannot be observed in non-dividing cells such as larval salivary gland cells with over-expression of the orbit. The induction of apoptosis is required for a proper function of mitotic checkpoint genes and p53. Our data should suggest that cells with abnormal microtubules deviated from proper range of the dynamic instability could be recognized during mitosis and could be eliminated by the apoptosis from proliferating tissues.

微管确实参与了许多细胞现象。研究表明，微管的动态不稳定性对某些细胞事件的调节特别重要。轨道/CLASP，一个保守的微管相关蛋白，富集在微管的正末端，在动态不稳定性的调节中起着至关重要的作用。果蝇轨道的功能丧失突变增加了微管的不稳定性相反，使用GAL 4-UAS系统在成虫盘中过表达该基因导致微管过度稳定，从而形成在间期的正常细胞中从未观察到的微管束。眼盘细胞在形态发生沟形成后，眼眶的过度表达使中期细胞聚集，这表明它激活了有丝分裂中的微管组装检查点。我们发现，过度表达最终诱导凋亡依赖于Caspase 3在有丝分裂后阶段的眼盘。基因的敲除也引起翅盘中半胱天冬酶3的诱导。这种类型的细胞凋亡不能在非分裂细胞中观察到，如具有眶过度表达的幼虫唾液腺细胞。细胞凋亡的诱导是有丝分裂检查点基因和p53的正常功能所必需的。我们的数据表明，异常微管偏离适当范围的动态不稳定性的细胞可以在有丝分裂过程中被识别，并可以被细胞凋亡从增殖组织中消除。

项目成果

Genetic link between ^-sarcoglycan and the Egfr signaling pathway.

^-肌聚糖和 Egfr 信号通路之间的遗传联系。

• 发表时间: 2006
• 期刊: Biochem. Biophys. Res. Commun. 348
• 作者: Hashimoto;R.;Yamaguchi;M.
• 通讯作者: M.

Genetic link between p-sarcoglycan and the Egfr signaling pathway.

p-肌聚糖和 Egfr 信号通路之间的遗传联系。

• 发表时间: 2006
• 期刊: Biochem.Biophys.Res.Commun. 348
• 作者: Hashimoto;R.;Yamaguchi;M.
• 通讯作者: M.

Antagonistic regulation of the Drosophila PCNA gene promoter by DREF and Cut

• DOI: 10.1111/j.1365-2443.2006.00956.x
• 发表时间: 2006-05-01
• 期刊: GENES TO CELLS
• 影响因子: 2.1
• 作者: Seto, H;Hayashi, Y;Yamaguchi, M
• 通讯作者: Yamaguchi, M

Transcriptional regulation of the Drosophila rfc1 gene by the DRE-DREF pathway

• DOI: 10.1111/j.1742-4658.2007.05730.x
• 发表时间: 2007-04-01
• 期刊: FEBS JOURNAL
• 影响因子: 5.4
• 作者: Tsuchiya, Akihiro;Inoue, Yoshihiro H.;Yamaguchi, Masamitsu
• 通讯作者: Yamaguchi, Masamitsu

Microtubule bundle formation and cell death induced by the human Orbit/CLASP N-terminal fragment.

人 Orbit/CLASP N 末端片段诱导微管束形成和细胞死亡。

• 发表时间: 2005
• 期刊: Cell Struct.Funct. 20
• 作者: Aonuma;M.;Miyamoto;M.;Inoue;Y.H.;et al.
• 通讯作者: et al.