DICER1与AR通过相分离调控染色质重塑在附睾抗菌中的分子机制研究

Our previous study indicated that DICER1 played a critical role in antimicrobial ability of epididymis and male fertility by regulating β-defensin gene transcription. Further exploration showed that β-defensin genes were modulated by DICER1 with AR together and chromatin remodeling was involved in too. Recent research suggested that there were strong links between phase separation, chromatin remodeling and transcription. Insterestingly, there are unique internal disorder regions in both DICER1 protein and AR protein which is the typical domain of phase separation protein. These suggested that DICER1 and AR might control transcription via phase separation and chromatin remodeling. This project will use conditional Dicer1 knockout mouse model combining with CO-IP, ChIP, nucleosome-scanning, 3C and FRAP technologies to verify the interation between DICER1 and AR. We want to explain this new biological function with novel insight mechanisms through phase separation and chromatin structure which DICER1 and AR were positioning on, relating and recruiting each other to co-regulate the mouse epididymal genes. Further more, we want to check that the mechanism has pervasive theoretical meaning and value...The findings of this new investigation will provide solid base for describing the mechanisms of antibacterial activity of epididymis, and provide new ideas for the clinical treatment of infection. Meanwhile, these investigations will also be valuable for future studies on DICER1 in other organs and other biological events.

我们的前期研究表明DICER1能调控β-defensin基因转录从而影响附睾抗菌和雄性生育。进一步的探索发现DICER1与转录因子AR能共同调控基因表达且涉及染色质构象重塑。有研究表明蛋白相分离与染色质重塑和基因转录存在密切关联，而DICER1和AR蛋白中均存在相分离蛋白的内部无序区，提示DICER1与AR可能通过相分离调控染色质重塑从而介导基因转录。本课题将利用条件性敲除Dicer1的小鼠模型，通过CO-IP、ChIP、核小体排列分析、3C、FRAP等技术探究DICER1与AR的相互作用，从相分离和染色质构象的角度阐释DICER1与AR共同调控附睾基因转录的新功能，新机制，并寻找该机制的普遍意义。.本课题的研究结果不但为理解附睾抗菌的分子机制提供理论支持，为临床治疗其他器官感染提供新的思路，也可以为其他组织器官和生物学过程中的基因转录调控研究提供重要的参考意义。

我们的前期研究表明DICER1能调控β-defensin基因转录从而影响附睾抗菌和雄性生育，且DICER1与转录因子AR能共同调控基因表达，其中涉及染色质构象重塑。进一步的研究发现，DICER1与AR调控的染色质构想重塑并不是通过蛋白质相分离实现，而是通过DNA相分离实现，从DNA相分离和染色质构象的角度阐释DICER1与AR共同调控附睾基因转录的新功能，新机制。.生物大分子的相分离现象普遍存在。本课题的研究结果不但为理解附睾抗菌的分子机制提供理论支持，为临床治疗其他器官感染提供新的思路，也可以为其他组织器官和生物学过程中的基因转录调控研究提供重要的参考意义。