RBM15结合促进RNA内含子或外显子滞留  

RBM15 Promotes Intron or Exon Retention

作　　者：王茹 柯岩[3] 韦荣飞 王新禹 李栋 WANG Ru;KE Yan;WEI Rong-Fei;WANG Xin-Yu;LI Dong(Division of Life Sciences and Medicine,University of Science and Technology of China,Hefei 230027,China;National Laboratory of Biomacromolecules,CAS Center for Excellence in Biomacromolecules,Institute of Biophysics,Chinese Academy of Sciences,Beijing100101,China;Arthritis Clinic and Research Center,Peking University Peopleʼs Hospital,Beijing 100044,China;College of Life Sciences,University of Chinese Academy of Sciences,Beijing100049,China)

机构地区：[1]中国科学技术大学生命科学与医学部生命科学学院,合肥230027 [2]中国科学院生物物理研究所,中国科学院科教融合生物大分子卓越创新中心,生物大分子国家重点实验室,北京100101 [3]北京大学人民医院骨关节科,北京100044 [4]中国科学院大学生命科学学院,北京100049

出　　处：《生物化学与生物物理进展》2021年第6期667-676,共10页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金(31771440,31770930)资助项目。

摘　　要：RBM15是一种RNA结合蛋白,参与到RNA的m6A修饰及可变剪接调控中.然而,RBM15在转录组水平如何调控可变剪接尚不清楚.本研究应用超分辨率荧光显微镜技术发现,RBM15在细胞核中形成斑点状结构,且与核斑有密切接触或完全定位于核斑中.核斑为细胞核中无膜细胞器,富含多种剪接因子,这提示RBM15可能参与到可变剪接的调控过程中.为了确定RBM15能否及如何调控可变剪接,我们利用siRNA敲低RBM15,并对敲低RBM15和野生型细胞进行二代测序.结果显示,敲低RBM15能够引起1111个转录本中的1279个可变剪接事件的变化.与已发表的RBM15-CLIP数据进行比较分析,我们发现,这1111个转录本中,有191个能够与RBM15结合,提示这191个转录本可能为RBM15调控的直接靶标.进一步的分析表明,RBM15结合能够促进这191个转录本中的121个发生内含子或外显子的滞留.该研究揭示了RBM15在转录组水平调控可变剪接的规律.RBM15 is an RNA binding protein that is known of involving in the m6 A modification and the regulation of alternative splicing(AS).However,how RBM15 regulates AS is currently unclear.Here,using super-resolution microscopy,we found that RBM15 forms puncta structures that closely contact with or even embedded in the nuclear speckles.Nuclear speckles are enriched in splicing factors,which implies that RMB15 might be involved in RNA AS.To determine whether and how RBM15 regulates AS,we knocked down RBM15(RBM15-KD)using si RNA and performed RNA-seq for wildtype(WT)cells and RBM15-KD cells.We analyzed the RNA-seq of WT and RBM15-KD cells.We show that RBM15-KD cause 1279 differential AS events in 1111 transcripts.After comparing to public RBM15-CLIP data,we identify that 191 out of 1111 transcripts are directly bound by RMB15,indicating that these 191 transcripts are probably direct targets of RBM15.Moreover,RBM15 promotes the retention of the adjacent regions proximal to its binding sites in 121 out of the 191 transcripts.This study reveals that how RBM15 regulates AS on a transcriptomic level.

关 键 词：RBM15 可变剪接 核斑 

分 类 号：Q7[生物学—分子生物学] Q219