Identification of Targets of CUG-BP, Elav-Like Family Member 1 (CELF1) Regulation in Embryonic Heart Muscle.
Identification of Targets of CUG-BP, Elav-Like Family Member 1 (CELF1) Regulation in Embryonic Heart Muscle.
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DOI：
10.1371/journal.pone.0149061
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Ladd AN
中科院分区：
文献类型：
--
作者：
Blech-Hermoni Y;Dasgupta T;Coram RJ;Ladd AN
CUG-BP，Elav-like家族成员1（CELF 1）是高度保守的RNA结合蛋白，其调节前mRNA选择性剪接、多腺苷酸化、mRNA稳定性和翻译。在心脏中，CELF 1在心肌中表达，其水平在发育过程中受到严格调控。CELF 1水平在胚胎发生期间在心脏中达到峰值，并且CELF 1在成人心脏中的异常上调与强直性肌营养不良1型以及糖尿病性心肌病的心脏发病机制有关。心肌中CELF活性的抑制或CELF 1的过度表达导致转基因小鼠的心肌病。尽管如此，CELF 1调节的许多心脏靶点仍然未知。在这项研究中，为了确定CELF 1的心脏靶点，我们对来自胚胎第8天的鸡心脏的CELF 1进行了交联免疫沉淀（CLIP）。我们确定了MYH 7 B中一个以前未注释的外显子作为CELF 1介导调控的新靶点。我们证明，在原代鸡胚心肌细胞中CELF 1的敲低导致该外显子的包含增加和MYH 7 B水平降低。我们还研究了在已发表的RNA-seq数据集中CELF 1敲低后原代胚胎心肌细胞转录组的总体变化。通路和网络分析确定了CELF 1与细胞周期和翻译调控之间的强关联。重要的调节蛋白，包括RNA结合蛋白和心脏转录因子，受到CELF 1丢失的影响。总之，这些数据表明CELF 1是心肌细胞基因表达的关键调节因子。
CUG-BP, Elav-like family member 1 (CELF1) is a highly conserved RNA binding protein that regulates pre-mRNA alternative splicing, polyadenylation, mRNA stability, and translation. In the heart, CELF1 is expressed in the myocardium, where its levels are tightly regulated during development. CELF1 levels peak in the heart during embryogenesis, and aberrant up-regulation of CELF1 in the adult heart has been implicated in cardiac pathogenesis in myotonic dystrophy type 1, as well as in diabetic cardiomyopathy. Either inhibition of CELF activity or over-expression of CELF1 in heart muscle causes cardiomyopathy in transgenic mice. Nonetheless, many of the cardiac targets of CELF1 regulation remain unknown. In this study, to identify cardiac targets of CELF1 we performed cross-linking immunoprecipitation (CLIP) for CELF1 from embryonic day 8 chicken hearts. We identified a previously unannotated exon in MYH7B as a novel target of CELF1-mediated regulation. We demonstrated that knockdown of CELF1 in primary chicken embryonic cardiomyocytes leads to increased inclusion of this exon and decreased MYH7B levels. We also investigated global changes in the transcriptome of primary embryonic cardiomyocytes following CELF1 knockdown in a published RNA-seq dataset. Pathway and network analyses identified strong associations between CELF1 and regulation of cell cycle and translation. Important regulatory proteins, including both RNA binding proteins and a cardiac transcription factor, were affected by loss of CELF1. Together, these data suggest that CELF1 is a key regulator of cardiomyocyte gene expression.