Zhang Eric E, Liu Andrew C, Hirota Tsuyoshi, Miraglia Loren J, Welch Genevieve, Pongsawakul Pagkapol Y, Liu Xianzhong, Atwood Ann, Huss Jon W, Janes Jeff, Su Andrew I, Hogenesch John B, Kay Steve A
Genomics Institute of the Novartis Research Foundation, San Diego, CA 92121, USA.
Cell. 2009 Oct 2;139(1):199-210. doi: 10.1016/j.cell.2009.08.031. Epub 2009 Sep 17.
Two decades of research identified more than a dozen clock genes and defined a biochemical feedback mechanism of circadian oscillator function. To identify additional clock genes and modifiers, we conducted a genome-wide small interfering RNA screen in a human cellular clock model. Knockdown of nearly 1000 genes reduced rhythm amplitude. Potent effects on period length or increased amplitude were less frequent; we found hundreds of these and confirmed them in secondary screens. Characterization of a subset of these genes demonstrated a dosage-dependent effect on oscillator function. Protein interaction network analysis showed that dozens of gene products directly or indirectly associate with known clock components. Pathway analysis revealed these genes are overrepresented for components of insulin and hedgehog signaling, the cell cycle, and the folate metabolism. Coupled with data showing many of these pathways are clock regulated, we conclude the clock is interconnected with many aspects of cellular function.
二十年的研究确定了十几种生物钟基因，并定义了昼夜节律振荡器功能的生化反馈机制。为了识别更多的生物钟基因和调节因子，我们在人类细胞生物钟模型中进行了全基因组小干扰RNA筛选。敲低近1000个基因会降低节律幅度。对周期长度有显著影响或增加幅度的情况则较少见；我们发现了数百个这样的基因，并在二次筛选中进行了验证。对这些基因的一个子集进行表征，结果表明其对振荡器功能具有剂量依赖性效应。蛋白质相互作用网络分析表明，数十种基因产物直接或间接与已知的生物钟成分相关联。通路分析显示，这些基因在胰岛素和刺猬信号通路、细胞周期以及叶酸代谢的成分中占比过高。再加上数据表明这些通路中的许多都受生物钟调节，我们得出结论，生物钟与细胞功能的许多方面相互关联。
