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Protein modification systems controlling/integrating entraintnznt, oscillation and output machinery of circadian clock

控制/整合生物钟的夹带、振荡和输出机制的蛋白质修饰系统

基本信息

批准号：
18500300
负责人：
TAMARU Teruya
金额：
$ 2.52万
依托单位：
Toho University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18500300/
关键词：
circadian clock ubiouitin phosphorylation transcription factor acetvlation protein kinase BMAL1 transcription nuclear entry pretein kinase reset

项目摘要

Control of the stability of protein is thought to involved in various biological processes, such as cell cycle and circadian clock. MG132, a reversible inhibitor for proteolysis of ubiquitinated (Ub-) proteins; this might be due to inhibition of BMAL1 degradation via Ub-proteasome. I showed that endogenous Ub-BMAL1 accumulated with MG132 during dexamethazone (Dex) -pulse treatment induced cellular clock synchronization/resetting of NIH-3T3 cells. Moreover, BMAL1 ubiquitination was suppressed by U0126, a phosphorylation inhibitor via ERK pathway. Inhibition of Ub-Proteasome during Dex-pulse significantly reduced circadian clock proteins during circadian cycle and damped out-put circadian rhythm of luciferase-reporting Per2 gene expression (Per2-luc rhythm). BMAL1 contains a conserved ERK phosphorylation site (T527) in its PEST-like sequence, which is thought to involved in the controlling protein stability. Ectopic expressed BMAL1 mutants, which lack PEST-like sequence (ΔPEST) or ERK … More phosphorylation site (T527A) , in BMAL1-deficient mouse. Embryonic fibroblasts MEFs) during clock-resetting resulted in dramatically reduced Ub-BMAL1 level in T527A and slightly higher level in ΔPEST. This strongly suggest that ERK phosphorylation in BMAL1-PEST is essential for the ubiquitination-controlled clock-resetting and naive controlled Ub-BMAL1 accumulation of wild type BMAL1 during clock-resetting is Unpaired. In BMAL1-ΔPEST. Transient transfection of GFP-BMAL1/PEST protein, which is expected to perturb ERK phosphorylation mediated ubiquitination during Dex-pulse resulted in reduced level of Ub-BMAL1 accumulation and dramatically damped Per2-luc rhythm. I propose that controlling stability of BMAL1 via ERK phohorylation dependent ubiquitination in its PEST-like sequence have essential roles during resetting/generating circadian clock oscillation.With collaboration with UCI team (Dept. Pharmacology, USA), I also demonstrated that essential role of circadian BMAL1 acetylation in mammalian clock system (Nature 2007). Less

蛋白质稳定性的控制被认为涉及到各种生物过程，如细胞周期和生物钟。MG132，泛素化(Ub-)蛋白降解的可逆抑制剂；这可能是由于抑制了Ub-蛋白酶体对BMAL1的降解。我发现在地塞米松(Dex)脉冲处理NIH-3T3细胞过程中，内源性Ub-BMAL1与MG132积聚，诱导NIH-3T3细胞时钟同步/重置。此外，BMAL1泛素化被ERK途径的磷酸化抑制剂U0126抑制。在Dex脉冲期间抑制Ub蛋白酶体显著减少了昼夜节律中的生物钟蛋白，并抑制了报告荧光素酶PER2基因表达的输出昼夜节律(PER2-Luc节律)。BMal1在其类似PEST的序列中含有一个保守的ERK磷酸化位点(T527)，被认为参与了控制蛋白质稳定性的过程。异位表达的BMAL1突变体，缺乏类似害虫的序列(ΔPEST)或ERK…在BMAL1缺陷小鼠中，有更多的磷酸化位点(T527A)。在时钟重置期间，胚胎成纤维细胞(MEF)在T527A中的Ub-BMAL1水平显著降低，而在ΔPEST中的水平略高。这表明BMAL1-PEST中ERK的磷酸化对于泛素化控制的时钟重置是必不可少的，而野生型BMAL1在时钟重置过程中天真控制的Ub-BMAL1积累是不成对的。在BMAL1-Δ害虫中。瞬时转染GFP-BMAL1/PEST蛋白，有望干扰ERK磷酸化介导的泛素化，导致Ub-BMAL1积聚水平降低，并显著抑制PER2-Luc节律。我认为，通过ERK磷酸化依赖的泛素化来控制BMAL1的稳定性，在其PEST样序列中，在重置/产生昼夜时钟振荡过程中起着至关重要的作用。我还展示了生物钟BMAL1乙酰化在哺乳动物时钟系统中的重要作用(自然，2007)。较少

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Role of post-translational modification of BMAL1 in mammalian circadian clock

BMAL1 翻译后修饰在哺乳动物生物钟中的作用

DOI：
发表时间：
2007
期刊：
影响因子：
0
作者：
Tamaru;T.;Takamatsu;K
通讯作者：
K

Circadian rhythmic CLOCK : BMAL1 kinase PFK controls mammalian clock function

昼夜节律时钟：BMAL1 激酶 PFK 控制哺乳动物时钟功能

DOI：
发表时间：
2008
期刊：
影响因子：
0
作者：
Tamaru;T.;Takamatsu;K
通讯作者：
K

「中枢時計と末梢時計:時計遺伝子研究の最前線」Bmall遺伝子と概日リズム形成

《中枢时钟与外周时钟：时钟基因研究的前沿》Bmall基因与昼夜节律形成

DOI：
发表时间：
2007
期刊：
Clinical Neuroscience 25
影响因子：
0
作者：
Hirayama J.;Sahaz S.;Grimaldi B.;Tamara T.;Takamatsu K.;Nakahata Y.;Sassone-Corsi P.;田丸輝也
通讯作者：
田丸輝也

日周期変動性CLOCK:BMAL1キナーゼPFKによる哺乳類体内時計の制御

昼夜节律时钟：BMAL1 激酶 PFK 对哺乳动物生物钟的控制