转录因子SOX2与长链非编码RNA在基因组调控中的相互作用机制研究

项目名称： 转录因子SOX2与长链非编码RNA在基因组调控中的相互作用机制研究

项目编号： No.31501058

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 遗传学与生物信息学、细胞生物学

项目作者： 侯琳琳

作者单位： 中国科学院广州生物医药与健康研究院

项目金额： 20万元

中文摘要： SOX2是SOX转录因子家族的成员，它在早期胚胎发育、神经分化以及癌症发生等过程中起到关键作用。最近研究发现长链非编码RNA以SOX2依赖的方式调控基因组进而决定干细胞命运，但它们在该过程中的相互作用机制至今仍不清楚。本课题组通过凝胶迁移实验发现SOX2可通过其DNA结合结构域－HMG结构域直接结合RNA进而影响其对DNA的结合。本项目拟利用凝胶迁移实验等技术检测SOX2的RNA结合特性；结合蛋白结晶和等温滴定量热法等技术构建SOX2的RNA结合模型；从人脐静脉内皮细胞中鉴定与SOX2结合的长链非编码RNA并通过荧光能量共振转移实验等手段检测它们对SOX2结合DNA序列的影响，从而阐明SOX2与长链非编码RNA在基因组调控中的相互作用机制。本研究将有助于理解其他SOX转录因子和长链非编码RNA的关系，深化认识细胞命运的决定机制，为相关疾病的治疗提供新的方向。

中文关键词： 转录因子SOX2；HMG结构域；长链非编码RNA；相互作用机制；基因组调控

英文摘要： SOX2 is a member of SOX (SRY-related high-mobility-group box) transcription factor family. It is a vital player in diverse processes, including early embryonic development, neural differentiation, and carcinogenesis. It was recently found that long noncoding RNAs (lncRNAs) play key roles in genome regulation and stem cell fate determination in a SOX2-dependent way. However, the cooperation mechanism between them are still unrevealed. Our data obtained through electrophoretic mobility shift assays (EMSA) show that SOX2 directly binds to RNA via its DNA-binding domain- HMG box, leading to influences on its DNA binding property. In this research, we will first test whether RNAs are bound by SOX2 in a sequence-specific or structure-specific manner through EMSA and fluorescence resonance energy transfer (FRET); next, in combine with crystallization trail and isothermal titration calorimetry (ITC), we will construct the model of interaction between SOX2 and RNA; then, SOX2-interacted lncRNAs will be ascertained in human umbilical vein endothelial cells (HUVECs) and their effects on DNA binding by SOX2 will be identified through EMSA and FRET. This work will contribute to understand the relations between other SOX transcription factors and lncRNAs and mechanism of cell fate determination, and support a new direction for related diseases treatments.

英文关键词： transcription factor SOX2;HMG domain;long noncoding RNA;interplay mechanism;genome regulation