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Molecular Biology of the Biological Clock : From Gene to Behavior

生物钟的分子生物学：从基因到行为

基本信息

批准号：
11470018
负责人：
OKAMURA Hitoshi
金额：
$ 9.66万
依托单位：
Kobe University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11470018/
关键词：
mPer1 clock genes dbp cry e4bp4 circadian rhythm transgenic mice suprachiasmatic nucleus 哺乳類時計遺伝子サーカディアンリズム Cry 単一細胞のリズム

项目摘要

In the eukaryotic circadian model systems, translocation of the oscillatory gene products into the nucleus is a key step for generation of a 24 hour cycle of the biological clock. We have examined nuclear import of clock proteins of the mammalian period gene family and the effect of serum shock, which induces a synchronous clock in cultured cells. We examined the nuclear import of mPER proteins in COS7 cells and found that nuclear translocation of mPER1 and mPER2 involves physical interactions with mPER3. This indicates that nuclear translocation of mPER1 also can occur under physiological conditions in the absence of CRY proteins.Recently we demonstrated that an accessory transcription loop exists helping to the core clock feedback loop. Transcript levels of DBP, a member of the PAR leucine zipper transcription factor family, exhibit a robust rhythm in the SCN.We report that DBP is able to activate the promoter of a putative clock oscillating gene, mPer1, by directly binding to the mPer1 promoter. DBP and CLOCK-BMAL1 cooperatively activate the mPer1 promoter. On the other hand, dbp transcription is activated by CLOCK-BMAL1 through E-boxes and inhibited by the mPER and mCRY proteins, as is the case for mPer1. Antiphase circadian regulated E4BP4, another member of leucine zipper transcription factor without PAR domain, antagonistically suppresses mPer1 transcription. Thus, a clock-controlled dbp and e4bp4 genes may play an important role in the central clock oscillation.Using transgenic mice carrying the mPer1 promoter fused to the luciferase (mPer1-luc) gene, we recently succeeded to monitor luciferase-mediated bioluminescence with a day-night variation in the SCN in brain slices and in living animals. We can record for several days oscillating photon emission with a periodicity and phase that accurately mirrored native mPer1 mRNA expression. The real-time optical imaging of gene expression will be a new powerful tool to study mammalian brain function.

在真核生物昼夜节律模型系统中，振荡基因产物易位到细胞核中是产生24小时周期生物钟的关键步骤。我们已经研究了哺乳动物周期基因家族的时钟蛋白的核输入和血清休克的影响，其在培养的细胞中诱导同步时钟。我们研究了COS 7细胞中mPER蛋白的核输入，发现mPER 1和mPER 2的核转位涉及与mPER 3的物理相互作用。这表明mPER 1在没有CRY蛋白的生理条件下也可以发生核转位。最近我们证明了一个辅助转录环的存在，它帮助核心时钟反馈环。转录水平的DBP，PAR亮氨酸拉链转录因子家族的成员，表现出强大的节奏在SCN。我们报告，DBP能够激活一个假定的时钟振荡基因，mPer 1的启动子，通过直接结合到mPer 1启动子。DBP和CLOCK-BMAL 1协同激活mPer 1启动子。另一方面，DBP转录被CLOCK-BMAL 1通过E-box激活，并被mPER和mCRY蛋白抑制，与mPer 1的情况相同。反相昼夜节律调节的E4 BP 4，另一个成员的亮氨酸拉链转录因子没有PAR结构域，拮抗性抑制mPer 1的转录。因此，一个时钟控制的DBP和e4 bp 4基因可能在中央时钟oscillation.Using转基因小鼠携带mPer 1启动子融合的荧光素酶（mPer 1-luc）基因，我们最近成功地监测在SCN在脑切片和活体动物的昼夜变化的脱氢酶介导的生物发光。我们可以记录几天振荡光子发射的周期和相位，准确反映了天然mPer 1 mRNA的表达。基因表达的实时光学成像将成为研究哺乳动物脑功能的新的有力工具。

项目成果

期刊论文数量（76）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Takumi T, Nagamine Y, Miyake S, Matsubara C, Taguchi K, Takekida S, Sakakida Y, Nishikawa K, Kishimoto T, Niwa S, Okumura K, Okamura H: "A mammalian orthologue of Drosophila timeless, highly expressed in SCN and retina, forms a complex with mPER1"Genes Ce

Takumi T、Nagamine Y、Miyake S、Matsubara C、Taguchi K、Takekida S、Sakakida Y、Nishikawa K、Kishimoto T、Niwa S、Okumura K、Okamura H：“果蝇永恒的哺乳动物直系同源物，在 SCN 和视网膜中高度表达