Gene structure, expression, and DNA methylation characteristics of sea cucumber cyclin B gene during aestivation
Gene structure, expression, and DNA methylation characteristics of sea cucumber cyclin B gene during aestivation
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海参cyclin B基因夏眠期间的基因结构、表达及DNA甲基化特征
DOI：
10.1016/j.gene.2016.09.006
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发表时间：
2016
期刊：
影响因子：
3.5
通讯作者：
Storey Kenneth B.
中科院分区：
文献类型：
--
作者：
Zhu Aijun;Chen Muyan;Zhang Xiumei;Storey Kenneth B.
刺参是研究海洋无脊椎动物环境诱导消亡的良好模型。夏眠的核心要求之一是抑制耗能的细胞过程，如细胞周期进程。本研究确定了细胞周期调节因子Cyclin B的基因结构，并检测了该基因在每年的夏眠-唤醒周期的三个阶段的表达水平。此外，还分析了细胞周期蛋白B在海参非去雄阶段和深去巢阶段的DNA甲基化特征。我们发现环素B启动子含有一个CpG岛、三个CCAAT盒和三个细胞周期基因同源区(CHR)。应用定量逆转录聚合酶链式反应(QRT-PCR)分析表明，与未去巢相比，深去巢期间肠道和纵肌中细胞周期蛋白B的转录水平均显著降低，在去巢后恢复到基础水平。对环素Bcore启动子的甲基化分析表明，其甲基化水平在未消亡阶段和深度消退阶段之间存在显著差异(p<；P<0.05)，并且发现核心启动子的甲基化水平与Cyclin B转录本的表达呈正相关。我们的研究结果表明，细胞周期进程在退缩过程中可能被可逆地阻止，这表明细胞周期蛋白B表达水平的变化，我们认为DNA甲基化是参与细胞周期蛋白B转录变异的调节机制之一。
The sea cucumber, Apostichopus japonicus, is a good model for studying environmentally-induced aestivation by a marine invertebrate. One of the central requirements of aestivation is the repression of energy-expensive cellular processes such as cell cycle progression. The present study identified the gene structure of the cell cycle regulator,cyclin B, and detected the expression levels of this gene over three stages of the annual aestivation-arousal cycle. Furthermore, the DNA methylation characteristics ofcyclin Bwere analyzed in non-aestivation and deep-aestivation stages of sea cucumbers. We found that thecyclin Bpromoter contains a CpG island, three CCAAT-boxes and three cell cycle gene homology regions (CHRs). Application of qRT-PCR analysis showed significant downregulation ofcyclin Btranscript levels during deep-aestivation in comparison with non-aestivation in both intestine and longitudinal muscle, and these returned to basal levels after arousal from aestivation. Methylation analysis of thecyclin Bcore promoter revealed that its methylation level showed significant differences between non-aestivation and deep-aestivation stages (p< 0.05) and interestingly, a positive correlation betweenCyclin Btranscripts expression and methylation levels of the core promoter was also observed. Our findings suggest that cell cycle progression may be reversibly arrested during aestivation as indicated by the changes incyclin Bexpression levels and we propose that DNA methylation is one of the regulatory mechanisms involved incyclin Btranscriptional variation.