半滑舌鳎dmrt1基因转录因子和甲基化调控的互作机制

Chinese tongue sole is an important economic marine fish species in China. Due to the large difference in male and female growth and the high ratio of male in production, it has become a main bottleneck affecting the tongue sole industry. Previous studies found that the sex of Chinese tongue sole is determined by both genetic and environmental factors, and dmrt1 is the sex determination gene. In this study, we will focus on the regulatory mechanism of sex determination gene from the views of transcription factor and epigenetics in Chinese tongue sole. We firstly investigate the transcriptional regulation activity of transcription factors (sox5 and c/ebpα) to elucidate its regulation mechanism on dmrt1 expression. Secondly we explicate the relationship between the specific hypermethylation region and dmrt1 expression in male and pseudomale fish, and to reveal the functional transformation of methylation regulation in the promoter region. Thirdly we analyze the effect of methylation on the regulation activity of sox5 and c/ebpα, and to clarify the interaction mechanism of transcription factor regulation and methylation regulation on dmrt1. The results of this study will provide a new idea and approach for the sex control technology of Chinese tongue sole.

半滑舌鳎是我国特有的名贵经济海水鱼类，由于其雌雄性生长差异巨大且雄性比例较高，已成为限制半滑舌鳎养殖产业发展的主要瓶颈。前期研究发现半滑舌鳎性别决定受遗传和环境共同作用，且具有性别决定基因（dmrt1）。因此，本项目拟以半滑舌鳎为研究载体，聚焦其性别决定基因，从转录因子和表观遗传两个水平，分析转录因子（sox5和c/ebpα）对dmrt1启动子的转录调控活性，阐明其调控dmrt1表达的功能机制；明确伪雄鱼和雌鱼特异高甲基化区域和dmrt1基因表达之间的关系及其时序性，揭示半滑舌鳎dmrt1基因启动子区域甲基化调控方式的功能转换；研究甲基化对sox5及c/ebpα与dmrt1启动子相互作用的影响，解析dmrt1基因转录因子调控和甲基化调控的互作机制，为半滑舌鳎性别控制技术的开发提供新思路和新途径。

半滑舌鳎是我国特有的名贵经济海水鱼类，由于其雌雄性生长差异巨大且雄性比例较高，已成为限制半滑舌鳎养殖产业发展的主要瓶颈。本项目以半滑舌鳎为研究对象，聚焦其雄性决定基因dmrt1，从转录因子和表观遗传两个水平，分析转录因子（c/ebpα和sox5）对dmrt1启动子的转录调控活性，阐明其调控dmrt1表达的功能机制。研究结果构建了高效的半滑舌鳎dmrt1重组启动子，为通过过表达技术研究半滑舌鳎性别相关基因提供了一个理想的内源启动子；对转录因子c/ebpα和sox5进行克隆、表达分析及功能验证，验证了dmrt1启动子雌/伪雄特异高甲基化区域，并验证其对dmrt1具有负调控作用，为半滑舌鳎性控育种提供了可靠的分子及表观靶标；初步验证了dmrt1基因转录因子调控和甲基化调控可能具有协同性，为分析半滑舌鳎雄性决定基因dmrt1的调控作用提供了新视角。上述成果发表SCI论文4篇，为半滑舌鳎性别控制技术的开发提供新思路和新途径，并为深入了解海水鱼类性别决定与分化的机制提供了重要的理论依据。