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卵母细胞特异性基因 Oog1 的启动子在转基因小鼠的雄性和雌性减数分裂生殖细胞中均有功能。

The promoter of the oocyte-specific gene, Oog1, functions in both male and female meiotic germ cells in transgenic mice.

机构信息

Laboratory of Reproductive Biology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.

出版信息

PLoS One. 2013 Jul 22;8(7):e68686. doi: 10.1371/journal.pone.0068686. Print 2013.

Abstract

Oog1 is an oocyte-specific gene whose expression is turned on in mouse oocytes at embryonic day (E) 15.5, concomitant with the time when most of the female germ cells stop proliferating and enter meiotic prophase. Here, we characterize the Oog1 promoter, and show that transgenic GFP reporter expression driven by the 2.7 kb and 3.9 kb regions upstream of the Oog1 transcription start site recapitulates the intrinsic Oog1 expression pattern. In addition, the 3.9 kb upstream region exhibits stronger transcriptional activity than does the 2.7 kb region, suggesting that regulatory functions might be conserved in the additional 1.2 kb region found within the 3.9 kb promoter. Interestingly, the longer promoter (3.9 kb) also showed strong activity in male germ cells, from late pachytene spermatocytes to elongated spermatids. This is likely due to the aberrant demethylation of two CpG sites in the proximal promoter region. One was highly methylated in the tissues in which GFP expression was suppressed, and another was completely demethylated only in Oog1pro3.9 male and female germ cells. These results suggest that aberrant demethylation of the proximal promoter region induced ectopic expression in male germ cells under the control of 3.9 kb Oog1 promoter. This is the first report indicating that sex-dependent gene expression is altered according to the length and the methylation status of the promoter region. Additionally, our results show that individual CpG sites are differentially methylated and play different roles in regulating promoter activity and gene transcription.

摘要

Oog1 是一种卵母细胞特异性基因,其表达在胚胎期(E)15.5 的小鼠卵母细胞中被激活,此时大多数雌性生殖细胞停止增殖并进入减数分裂前期。在这里,我们对 Oog1 启动子进行了特征描述,并表明由 Oog1 转录起始位点上游的 2.7 kb 和 3.9 kb 区域驱动的转基因 GFP 报告基因表达可再现内在的 Oog1 表达模式。此外,3.9 kb 上游区域比 2.7 kb 区域具有更强的转录活性,这表明在 3.9 kb 启动子内发现的另外 1.2 kb 区域中可能保守了调节功能。有趣的是,较长的启动子(3.9 kb)在雄性生殖细胞中也表现出很强的活性,从晚期粗线期精母细胞到伸长的精子细胞。这可能是由于近端启动子区域的两个 CpG 位点异常去甲基化所致。一个在 GFP 表达被抑制的组织中高度甲基化,另一个仅在 Oog1pro3.9 雄性和雌性生殖细胞中完全去甲基化。这些结果表明,近端启动子区域的异常去甲基化在 3.9 kb Oog1 启动子的控制下导致雄性生殖细胞中的异位表达。这是第一个表明性别依赖性基因表达根据启动子区域的长度和甲基化状态而改变的报告。此外,我们的结果表明,个别 CpG 位点被不同程度地甲基化,并在调节启动子活性和基因转录方面发挥不同的作用。

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