The RNA-binding profile of the splicing factor SRSF6 in immortalized human pancreatic β-cells.
The RNA-binding profile of the splicing factor SRSF6 in immortalized human pancreatic β-cells.
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剪接因子SRSF6在永生化人胰腺β细胞中的RNA结合谱。
DOI：
10.26508/lsa.202000825
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发表时间：
2021-03
影响因子：
4.4
通讯作者：
Eizirik DL
中科院分区：
文献类型：
--
作者：
Alvelos MI;Brüggemann M;Sutandy FR;Juan-Mateu J;Colli ML;Busch A;Lopes M;Castela Â;Aartsma-Rus A;König J;Zarnack K;Eizirik DL
RNA结合蛋白SRSF 6识别由GAA三联体组成的富含嘌呤的共有基序，其在人胰腺β细胞中的下调以位置依赖性方式影响选择性剪接。在胰腺β细胞中，剪接因子SRSF 6的表达受GLIS 3调节，GLIS 3是一种由糖尿病易感基因编码的转录因子。SRSF 6下调通过剪接β细胞功能和存活的中心基因的失调促进β细胞死亡，但SRSF 6如何靶向RNA仍然知之甚少。在这里，我们通过在基础条件下整合单个核苷酸分辨率UV交联和免疫沉淀（iCLIP）以及SRSF 6敲低后的RNA测序，定义了人胰腺β细胞系EndoC-βH1中的SRSF 6结合景观。我们在编码序列中检测到数千个SRSF 6结合位点。基序分析表明，SRSF 6特异性地识别由GAA三联体组成的富含嘌呤的一致基序，并且连续GAA三联体的数量与结合位点强度的增加相关。SRSF 6定位决定剪接命运。根据SRSF 6在β细胞功能中的作用，我们鉴定了几个糖尿病易感基因中受调控外显子上的SRSF 6结合位点。在原理证明中，易感基因LMO 7的剪接由反义寡核苷酸调节。我们目前的研究揭示了SRSF 6在永生化人胰腺β细胞中的剪接调控景观。
The RNA-binding protein SRSF6 recognizes a purine-rich consensus motif consisting of GAA triplets, and its downregulation in human pancreatic β-cells affects alternative splicing in a position-dependent manner. In pancreatic β-cells, the expression of the splicing factor SRSF6 is regulated by GLIS3, a transcription factor encoded by a diabetes susceptibility gene. SRSF6 down-regulation promotes β-cell demise through splicing dysregulation of central genes for β-cells function and survival, but how RNAs are targeted by SRSF6 remains poorly understood. Here, we define the SRSF6 binding landscape in the human pancreatic β-cell line EndoC-βH1 by integrating individual-nucleotide resolution UV cross-linking and immunoprecipitation (iCLIP) under basal conditions with RNA sequencing after SRSF6 knockdown. We detect thousands of SRSF6 bindings sites in coding sequences. Motif analyses suggest that SRSF6 specifically recognizes a purine-rich consensus motif consisting of GAA triplets and that the number of contiguous GAA triplets correlates with increasing binding site strength. The SRSF6 positioning determines the splicing fate. In line with its role in β-cell function, we identify SRSF6 binding sites on regulated exons in several diabetes susceptibility genes. In a proof-of-principle, the splicing of the susceptibility gene LMO7 is modulated by antisense oligonucleotides. Our present study unveils the splicing regulatory landscape of SRSF6 in immortalized human pancreatic β-cells.