Signal transduction mechanism of cardiac stretch-activated channels

心脏牵张激活通道的信号转导机制

基本信息

批准号：
10670634
负责人：
FURUKAWA Tetsushi
金额：
$ 2.37万
依托单位：
Akita University (1999)Tokyo Medical and Dental University (1998)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670634/
关键词：
Ion channel Chloride Stretch Yeast two-hybrid Protein-protein interaction Cell cycle p34ィイD1cdc2ィエD1 cyclin B MAP kinase 伸展刺激細胞骨格アポトーシス圧負荷

项目摘要

The cardiac myocytes are always under the influence of physical force, and ion channels regulated by stretch may play important physiological roles in the heart. This research project, therefore, was designed to clarify the signal transduction mechanism around the stretch-activated ion channels in the molecular level. Candidate molecules interacting with several stretch-activated chloride channels were screened from cardiac cDNA library using the yeast two-hybrid method. The carboxyl terminus of ClC-2 channel was found to interact with three molecules that are regulated during the course of cell cycle. In vitro kinase assay revealed that the carboxyl terminus of ClC-2 was phosphorylated by p34ィイD1cdc2ィエD1/cycin B and MAP kinase. ClC-2 currents expressed in Xenopus oocytes were inhibited by p34ィイD1cdc2ィエD1/cyclin B, and were augmented by olomoucine, a cyclin-dependent kinase inhibitor. The reverse transcription PCR technique showed that the mRNA level of ClC-2 was also regulated by cell cycle, and was up-regulated in the fetal and hypertrophied heart. These data showed that ClC-2 channel was regulated by cell cycle machinery in the level of transcription, protein phosphorylation, and channel function. These mechanism may be important to achieve drastic change of cell volume during the course of cell cycle.

心肌细胞始终受到物理力的影响，拉伸调节的离子通道可能在心脏中发挥重要的生理作用，因此，本研究项目旨在阐明心脏中拉伸激活的离子通道周围的信号转导机制。使用酵母双杂交方法从心脏 cDNA 文库中筛选出与几个拉伸激活的氯离子通道相互作用的候选分子，发现 ClC-2 通道的羧基末端与三个受调节的分子相互作用。体外激酶测定表明，ClC-2 的羧基末端被 p34D1/cycin B 磷酸化，并且爪蟾卵母细胞中表达的 ClC-2 电流被 p34 D1cdc2D1/cyclin B 抑制。奥洛穆辛（一种细胞周期蛋白依赖性激酶抑制剂）增强了这一作用。转录PCR技术表明ClC-2的mRNA水平也受到细胞周期的调节，并且在胎儿和肥大的心脏中表达上调。这些数据表明ClC-2通道在转录水平上受到细胞周期机制的调节。、蛋白质磷酸化和通道功能对于在细胞周期过程中实现细胞体积的剧烈变化可能很重要。