Enhanced CLIP Uncovers IMP Protein-RNA Targets in Human Pluripotent Stem Cells Important for Cell Adhesion and Survival.
Enhanced CLIP Uncovers IMP Protein-RNA Targets in Human Pluripotent Stem Cells Important for Cell Adhesion and Survival.
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DOI：
10.1016/j.celrep.2016.03.052
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发表时间：
2016-04-19
期刊：
影响因子：
8.8
通讯作者：
Yeo GW
中科院分区：
文献类型：
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作者：
Conway AE;Van Nostrand EL;Pratt GA;Aigner S;Wilbert ML;Sundararaman B;Freese P;Lambert NJ;Sathe S;Liang TY;Essex A;Landais S;Burge CB;Jones DL;Yeo GW
人类多能干细胞（hPSC）需要精确控制转录后RNA网络以维持增殖和存活。使用增强的UV交联和免疫沉淀（eCLIP），我们鉴定了hPSC中IMP/IGF 2BP家族RNA结合蛋白的RNA靶点。在广泛的区域和结合位点水平，IMP 1和IMP 2显示出与大量重叠的3′ UTR富集靶点的可重复结合。应用于重组全长IMP 1和IMP 2的RNA Bind-N-Seq揭示了富含CA的基序，其富含eCLIP定义的结合位点。我们观察到，在hPSC中IMP 1的损失重演IMP 1表型，包括细胞粘附减少和细胞死亡增加。对于细胞粘附，在hPSC中，我们发现IMP 1维持整合素mRNA的水平，特异性地调节ITGB 5的RNA稳定性。此外，我们表明IMP 1可以通过直接靶向BCL 2与hPSC存活相关。因此，全转录组结合谱识别了调节已充分表征的IMP 1作用的hPSC靶点。
Human pluripotent stem cells (hPSCs) require precise control of post-transcriptional RNA networks to maintain proliferation and survival. Using enhanced UV crosslinking and immunoprecipitation (eCLIP), we identify RNA targets of the IMP/IGF2BP family of RNA-binding proteins in hPSCs. At the broad region- and binding site-level IMP1 and IMP2 show reproducible binding to a large and overlapping set of 3′UTR-enriched targets. RNA Bind-N-Seq applied to recombinant full-length IMP1 and IMP2 reveals CA-rich motifs that are enriched in eCLIP-defined binding sites. We observe that IMP1 loss in hPSCs recapitulates IMP1 phenotypes, including a reduction in cell adhesion and an increase in cell death. For cell adhesion, in hPSCs we find IMP1 maintains levels of integrin mRNA, specifically regulating RNA stability of ITGB5. Additionally, we show IMP1 can be linked to hPSC survival via direct target BCL2. Thus, transcriptome-wide binding profiles identify hPSC targets modulating well-characterized IMP1 roles.