New role of zCRY and zPER2 as regulators of sub-cellular distributions of zCLOCK and zBMAL proteins
New role of zCRY and zPER2 as regulators of sub-cellular distributions of zCLOCK and zBMAL proteins
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DOI：
10.1093/nar/gkg174
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发表时间：
2003-02-01
影响因子：
14.9
通讯作者：
Todo, T
中科院分区：
文献类型：
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作者：
Hirayama, J;Fukuda, I;Todo, T
真核生物产生昼夜节律的核心振荡器由基于转录/翻译的自调节反馈回路组成，时钟基因产物通过该回路负向调节其自身表达。负调节因子PER和CRY的积累和核进入的控制被认为是这些循环的关键步骤。我们阐明了斑马鱼时钟相关蛋白及其在NIH3T3细胞中的亚细胞定位之间的相互作用。斑马鱼中存在6种CRYs，其中zCRY1a强烈抑制zCLOCK1: zbmal3介导的转录，而zCRY3则不。我们发现zCRY1a与zCLOCK1和zBMAL3相互作用，促进核积累，而zCRY3与两者都不相关，也不影响它们的亚细胞分布。我们克隆了zPer2 cDNA，发现该cDNA编码的蛋白产物具有中等转录抑制因子的作用。在我们的亚细胞定位研究中，我们还发现zPER2与zCLOCK1:zBMAL3异源二聚体相互作用，导致其细胞质保留。zCRY1a和zPER2明显对zCLOCK:zBMAL异源二聚体的亚细胞分布有相反的影响。我们推测，这种对亚细胞分布的相反调控与zCRY1a和zPER2不同的转录抑制能力有关。zCRY1a通过直接与细胞核中的zCLOCK:zBMAL异源二聚体相互作用而成为一种有效的转录抑制剂，而zPER2在细胞质中维持zCLOCK:zBMAL异源二聚体，从而导致反活化抑制。
The core oscillator that generates circadian rhythm in eukaryotes consists of transcription/translation-based autoregulatory feedback loops by which clock gene products negatively regulate their own expression. Control of the accumulation and nuclear entry of the negative regulators PER and CRY is believed to be a key step in these loops. We clarified the mutual interaction between zebrafish clock-related proteins and their sub-cellular localizations in NIH3T3 cells. Six CRYs exist in zebrafish, of which zCRY1a strongly represses zCLOCK1: zBMAL3-mediated transcription, but zCRY3 does not. We show that zCRY1a interacts with zCLOCK1 and zBMAL3, facilitating nuclear accumulation, whereas zCRY3 associates with neither one and does not influence their sub-cellular distributions. We cloned zPer2 cDNA and showed that the protein product encoded by the cDNA acts as a moderate transcriptional repressor. In our sub-cellular localization studies we also found that zPER2 interacts with the zCLOCK1:zBMAL3 heterodimer, causing its cytoplasmic retention. zCRY1a and zPER2 apparently have opposite effects on the sub-cellular distribution of zCLOCK:zBMAL heterodimer. We speculate that the opposite regulation of the sub-cellular distribution of this is associated with the different transcriptional repression abilities of zCRY1a and zPER2. zCRY1a acts as a potent transcriptional inhibitor by interacting directly with the zCLOCK:zBMAL heterodimer in the nucleus, whereas zPER2 maintains the zCLOCK:zBMAL heterodimer in the cytoplasm, resulting in transactivation repression.