Jiangsu Key Laboratory of Sericultural and Animal Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, China.
Key Laboratory of Silkworm and Mulberry Genetic Improvement, Ministry of Agriculture and Rural Affairs, Sericultural Scientific Research Center, Chinese Academy of Agricultural Sciences, Zhenjiang 212100, China.
Genes (Basel). 2024 Oct 21;15(10):1351. doi: 10.3390/genes15101351.
The Jun proto-oncogene (), also referred to as , is an integral component of the JNK signaling pathway, which is crucial for the formation and differentiation of spermatogonial stem cells (SSCs). Investigations into the transcriptional regulation of chicken can offer a molecular foundation for elucidating its mechanistic role in SSCs. In this study, we successfully cloned a 2000 bp upstream sequence of the transcription start site and constructed a series of pGL3 recombinant vectors containing promoters of varying lengths. We verified the promoter activity of the 2000 bp upstream sequence by assessing the fluorescence intensity of DF-1 and identified the promoter activities of different regions via dual-luciferase assays. The transcription of JUN and its promoter region spanning -700 to 0 bp was modulated by an activator of the JNK signaling pathway. Bioinformatics analysis revealed that this -700 to 0 bp region was highly conserved among avian species and predicted the presence of binding sites for Wilms tumor 1 () and CCAAT/enhancer binding protein alpha (). The JNK signaling pathway activator was found to upregulate the expression of these transcription factors in DF-1 cells. Through the deletion of binding sites and the overexpression of and , we demonstrated that inhibited the transcription of , while promoted it. In conclusion, the -700 to 0 bp region is the key region of the promoter, with inhibiting transcription. The results of the study not only provide ideas for exploring the regulatory mechanism of in chicken SSCs, but also lay an important foundation for the study of avian SSCs.
原癌基因 Jun(),也被称为 c-Jun,是 JNK 信号通路的一个组成部分,对于精原干细胞(SSCs)的形成和分化至关重要。研究鸡的转录调控可以为阐明其在 SSCs 中的机制作用提供分子基础。
在这项研究中,我们成功克隆了 2000 bp 转录起始位点上游的序列,并构建了一系列包含不同长度启动子的 pGL3 重组载体。我们通过评估 DF-1 的荧光强度来验证 2000 bp 上游序列的启动子活性,并通过双荧光素酶测定确定了不同区域的启动子活性。JUN 的转录及其跨越 -700 至 0 bp 的启动子区域受到 JNK 信号通路激活剂的调节。生物信息学分析表明,该 -700 至 0 bp 区域在禽类中高度保守,并预测存在 Wilms 肿瘤 1(WT1)和 CCAAT/增强子结合蛋白α(C/EBPα)的结合位点。JNK 信号通路激活剂被发现可上调 DF-1 细胞中这些转录因子的表达。通过删除结合位点和过表达 和 ,我们证明了 抑制了 的转录,而 则促进了它的转录。
综上所述,-700 至 0 bp 区域是 启动子的关键区域, 抑制 转录。该研究的结果不仅为探索鸡 SSCs 中 调控机制提供了思路,也为禽类 SSCs 的研究奠定了重要基础。