A serine cluster mediates BMAL1-dependent CLOCK phosphorylation and degradation.
A serine cluster mediates BMAL1-dependent CLOCK phosphorylation and degradation.
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DOI：
10.4161/cc.8.24.10273
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发表时间：
2009-12-15
期刊：
影响因子：
--
通讯作者：
Antoch MP
中科院分区：
文献类型：
--
作者：
Spengler ML;Kuropatwinski KK;Schumer M;Antoch MP
生物钟以精确、持续的节奏调节从基因表达到生物体行为的生物过程，该节奏是在单细胞水平上通过互锁的转录反馈环和核心时钟蛋白的翻译后修饰的协调功能产生的。CLOCK磷酸化调节转录活性、细胞定位和稳定性;然而，对所涉及的特定残基和酶知之甚少。我们已经确定了一个保守的丝氨酸簇，包括Ser 431，这是一个先决条件的磷酸化位点的产生BMAL依赖性磷酸引发的时钟和潜在的GSK-3磷酸化Ser 427。突变分析和蛋白质稳定性测定表明，该丝氨酸簇作为磷酸降解决定子发挥功能。通过使用GSK-3激活剂/抑制剂和激酶测定，我们证明GSK-3β通过增加CLOCK磷酸化/降解来调节degron位点，这与CLOCK应答启动子表达的增加相关。磷酸缺陷型CLOCK的稳定化延迟了同步化成纤维细胞的振荡阶段。这项研究开始了一个复杂的磷酸调控位点的表征，控制时钟，核心转录因子，是必不可少的昼夜节律行为的降解。
The circadian clock regulates biological processes from gene expression to organism behavior in a precise, sustained rhythm that is generated at the unicellular level by coordinated function of interlocked transcriptional feedback loops and post-translational modifications of core clock proteins. CLOCK phosphorylation regulates transcriptional activity, cellular localization and stability; however little is known about the specific residues and enzymes involved. We have identified a conserved cluster of serines that include, Ser431, which is a prerequisite phosphorylation site for the generation of BMAL dependent phospho-primed CLOCK and for the potential GSK-3 phosphorylation at Ser427. Mutational analysis and protein stability assays indicate that this serine cluster functions as a phospho-degron. Through the use of GSK-3 activators/inhibitors and kinase assays, we demonstrate that GSK-3β regulates the degron-site by increasing CLOCK phosphorylation/degradation, which correlates with an increase in the expression of CLOCK responsive promoters. Stabilization of phospho-deficient CLOCK delays the phase of oscillation in synchronized fibroblasts. This investigation begins the characterization of a complex phospho-regulatory site that controls the degradation of CLOCK, a core transcription factor that is essential for circadian behavior.
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DOI：
10.1073/pnas.0409897102
发表时间：
2005-03-01
影响因子：
11.1
作者：
Gorbacheva, VY;Kondratov, RV;Antoch, MP
通讯作者：
Antoch, MP
影响因子：
10.5
作者：
Kondratov, RV;Chernov, MV;Antoch, MP
通讯作者：
Antoch, MP
影响因子：
56.9
作者：
Balsalobre, A;Brown, SA;Schibler, U
通讯作者：
Schibler, U
影响因子：
5.3
作者：
Kwon, Ilmin;Lee, Jiwon;Lee, Kun Ho
通讯作者：
Lee, Kun Ho
DOI：
10.1073/pnas.0811410106
发表时间：
2008-12-30
影响因子：
11.1
作者：
Hirota, Tsuyoshi;Lewis, Warren G.;Kay, Steve A.
通讯作者：
Kay, Steve A.