果蝇RNA m^(6)A修饰通路因子的筛选  

Screening of Drosophila melanogaster RNA m^(6)A modification pathway factors

作　　者：高舒阳 陆厚光 王艳花 严冬 Shuyang Gao;Houguang Lu;Yanhua Wang;Dong Yan(State Key Laboratory of Genetic Engineering,School of Life Sciences,Fudan University,Shanghai 200438,China)

机构地区：[1]复旦大学生命科学学院,遗传工程国家重点实验室,上海200438

出　　处：《遗传》2025年第4期476-488,共13页Hereditas(Beijing)

基　　金：国家自然科学基金项目(编号:31970786,32270868)资助。

摘　　要：N^(6)-甲基腺嘌呤(N^(6)-methyladenosine,m^(6)A)是mRNA中含量最丰富的化学修饰之一,在动植物发育和各种生理病理过程中发挥关键性的作用。先前的研究已经发现了m^(6)A甲基转移酶复合体、去甲基化酶以及m^(6)A结合蛋白,但作为一条相对较新的RNA修饰通路,可能还存在新的m^(6)A修饰因子。为了探究m^(6)A修饰对组织和器官的影响,本研究在果蝇(Drosophila melanogaster)眼成虫盘中过表达m^(6)A结合蛋白基因Ythdc1。结果显示,过表达Ythdc1导致雄性异位表达Sxl蛋白,雌雄均出现粗糙眼表型,同时也激活了JNK和细胞凋亡通路。为了利用粗糙眼表型进行m^(6)A修饰因子的筛选,本研究进一步构建了过表达Ythdc1的稳定果蝇品系。通过对1,500多个果蝇RNAi品系的筛选,成功鉴定到多个可能参与m^(6)A修饰的抑制子和增强子。这些基因目前在m^(6)A中的研究较少,因此进一步对它们进行了验证和初步的机制探索。总之,本研究发现了m^(6)A修饰通路的多个潜在因子,丰富了对m^(6)A修饰通路调控网络的认知,为探索m^(6)A修饰通路新的调控机制提供了思路和方向。N^(6)-methyladenosine(m^(6)A),one of the most prevalent mRNA modifications,plays crucial roles during animal and plant development and in various physiological and pathological processes.Previous studies have characterized m^(6)A methyltransferase complexes,demethylases,and m^(6)A-binding proteins,but as a relatively new epitranscriptomic pathway,it is likely that new m^(6)A components remain to be discovered.To explore the effects of m^(6)A modification on tissues and organs,the m^(6)A reader Ythdc1 was overexpressed in Drosophila melanogaster eye imaginal discs.Our results showed that overexpression of Ythdc1 leads to ectopic expression of Sxl in males,the rough eye in both males and females,and the activation of JNK signaling and apoptotic pathway.In order to screen m^(6)A modifiers using the rough eye phenotype,a stable Drosophila strain overexpressing Ythdc1 was further constructed.By screening of more than 1,500 RNAi lines,several repressors and enhancers that may be involved in m^(6)A modification were successfully identified.These genes are less studied in m^(6)A pathway,and therefore we further verified them and conducted preliminary mechanistic analyses on them.In summary,this study identified multiple potential factors of the m^(6)A modification pathway,expanded our understanding of the m^(6)A modification network,and provided ideas and directions for exploring new regulatory mechanisms of this important pathway.

关 键 词：果蝇 N6-甲基腺嘌呤 Ythdc1 Sxl 修饰因子筛选 

分 类 号：Q78[生物学—分子生物学]