Identification of ESRP1 targets in the epidermis

The epithelial splicing regulatory proteins, ESRP1 and ESRP2 are essential for mammalian development through regulation of a global program of alternative splicing of genes involved in maintenance of epithelial cell function. To further inform our understanding of the molecular functions of ESRP1 we performed enhanced crosslinking immunoprecipitation coupled with high throughput sequencing (eCLIP) in epithelial cells of mouse epidermis. The genome-wide binding sites of ESRP1 were integrated with RNA-Seq analysis of alterations in splicing and total gene expression that result from epidermal ablation of Esrp1 and Esrp2. These studies demonstrated that ESRP1 functions in splicing regulation occur primarily through direct binding in a position-dependent manner to either promote exon inclusion or skipping. In addition, we also identified widespread binding of ESRP1 in 3’ and 5’ untranslated regions (UTRs) of genes involved in epithelial cell function, suggesting that its post-transcriptional functions extend beyond splicing regulation. Crosslinking immunoprecipitation of ESRP1 from mouse epidermis, for nucleotide level resolution of ESRP1:RNA interactions

上皮剪接调控蛋白（epithelial splicing regulatory proteins）ESRP1与ESRP2，可通过调控参与上皮细胞功能维持的基因的全局可变剪接程序，对哺乳动物发育发挥至关重要的作用。为进一步阐明ESRP1的分子功能，我们在小鼠表皮上皮细胞中开展了增强型交联免疫沉淀联合高通量测序（enhanced crosslinking immunoprecipitation coupled with high throughput sequencing，eCLIP）实验。我们将ESRP1的全基因组结合位点与RNA测序（RNA-Seq）分析结果进行整合，该分析旨在探究表皮敲除Esrp1与Esrp2后可变剪接及基因总表达水平的变化。本研究证实，ESRP1的剪接调控功能主要通过以位置依赖的方式直接结合靶标RNA来实现，进而促进外显子保留或外显子跳跃。此外，我们还在参与上皮细胞功能维持的基因的3'与5'非翻译区（untranslated regions，UTRs）中发现了ESRP1的广泛结合位点，这提示其转录后调控功能并不仅限于剪接调控。本数据集包含小鼠表皮来源的ESRP1交联免疫沉淀数据，用于解析ESRP1与RNA相互作用的核苷酸级分辨率特征。