Splicing Factor RBM20 Regulates Transcriptional Network of Titin Associated and Calcium Handling Genes in The Heart.
Splicing Factor RBM20 Regulates Transcriptional Network of Titin Associated and Calcium Handling Genes in The Heart.
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DOI：
10.7150/ijbs.24117
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发表时间：
2018
影响因子：
9.2
通讯作者：
Greaser ML
中科院分区：
文献类型：
--
作者：
Guo W;Zhu C;Yin Z;Wang Q;Sun M;Cao H;Greaser ML
RNA结合基序20（RBM 20）调控30多个基因的前体mRNA剪接，其中肌联蛋白是主要靶点。在RBM 20表达的情况下，肌联蛋白在胎儿阶段表达较大的同种型，而在成人阶段表达较小的同种型，这是由选择性剪接引起的，而在没有RBM 20的情况下，肌联蛋白在所有年龄段中仅表达较大的同种型。除了剪接调控外，RBM 20是否也调控基因表达尚不清楚。在本研究中，我们采用Rbm 20基因敲除大鼠，研究基因表达谱使用Affymbase表达阵列。我们在第1、20和49天比较了野生型与Rbm 20敲除。生物信息学分析表明，RBM 20在年轻时调控的基因表达量较少，在老年时调控的基因表达量较多，并且常见表达基因也有相同的变化趋势。GSEA提示上调基因与心力衰竭相关。我们检查了肌联蛋白结合伴侣。所有肌联蛋白的直接结合伴侣都上调，其表达增加与扩张型心肌病相关。特别是，我们发现涉及钙处理和肌肉收缩的基因被RBM 20改变。对单个心肌细胞的细胞内钙水平测量进一步证实了这些蛋白质的变化影响钙处理。用QPCR和蛋白质印迹法验证来自肌联蛋白结合配偶体和钙处理的选定基因。这些数据表明，RBM 20调节基因剪接以及基因表达。通过RBM 20改变基因表达影响蛋白质-蛋白质相互作用、钙释放，从而影响肌肉收缩。我们的研究结果首次报道了RBM 20影响心脏成熟的基因表达，并为RBM 20在心力衰竭进展中的作用提供了新的见解。
RNA binding motif 20 (RBM20) regulates pre-mRNA splicing of over thirty genes, among which titin is a major target. With RBM20 expression, titin expresses a larger isoform at fetal stage to a smaller isoform at adult resulting from alternative splicing, while, without RBM20, titin expresses exclusively a larger isoform throughout all ages. In addition to splicing regulation, it is unknown whether RBM20 also regulates gene expression. In this study, we employed Rbm20 knockout rats to investigate gene expression profile using Affymetrix expression array. We compared wild type to Rbm20 knockout at day1, 20 and 49. Bioinformatics analysis showed RBM20 regulates fewer genes expression at younger age and more at older age and commonly expressed genes have the same trends. GSEA indicated up-regulated genes are associated with heart failure. We examined titin binding partners. All titin direct binding partners are up-regulated and their increased expression is associated with dilated cardiomyopathy. Particularly, we found that genes involving calcium handling and muscle contraction are changed by RBM20. Intracellular calcium level measurement with individual cardiomyocytes further confirmed that changes of these proteins impact calcium handling. Selected genes from titin binding partners and calcium handling were validated with QPCR and western blotting. These data demonstrate that RBM20 regulates gene splicing as well as gene expression. Altered gene expression by RBM20 influences protein-protein interaction, calcium releasing and thus muscle contraction. Our results first reported gene expression impacted by RBM20 with heart maturation, and provided new insights into the role of RBM20 in the progression of heart failure.
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影响因子：
3.4
作者：
Györke, I;Hester, N;Györke, S
通讯作者：
Györke, S
影响因子：
--
作者：
Guo, Wei;Sun, Mingming
通讯作者：
Sun, Mingming
影响因子：
2.7
作者：
DHOOT, GK;HALES, MC;PERRY, SV
通讯作者：
PERRY, SV
影响因子：
20.1
作者：
Borbely, Attila;Falcao-Pires, Ines;Paulus, Walter J.
通讯作者：
Paulus, Walter J.
影响因子：
20.1
作者：
Ching, LL;Williams, AJ;Sitsapesan, R
通讯作者：
Sitsapesan, R