Dose-Dependent Regulation of Alternative Splicing by MBNL Proteins Reveals Biomarkers for Myotonic Dystrophy
Dose-Dependent Regulation of Alternative Splicing by MBNL Proteins Reveals Biomarkers for Myotonic Dystrophy
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DOI：
10.1371/journal.pgen.1006316
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发表时间：
2016-09-01
期刊：
影响因子：
4.5
通讯作者：
Berglund, J. Andrew
中科院分区：
文献类型：
--
作者：
Wagner, Stacey D.;Struck, Adam J.;Berglund, J. Andrew
选择性剪接是一个受调控的过程，导致特定mRNA和蛋白质亚型的表达。不同的剪接因子决定了每个同工异构体的相对丰度。在这里，我们关注MBNL1，一种在肌强直性营养不良疾病中被错误调节的剪接因子。通过在很宽的动态范围内改变细胞中MBNL1的浓度，我们发现不同的剪接事件需要不同数量的MBNL1来达到半最大反应，并且对MBNL1的反应或多或少是陡峭的。研究了MBNL1外显子5周围的基序，以评估顺式元件如何介导MBNL1剂量依赖性剪接反应。我们开发了一个框架，仅通过剪接反应来估计MBNL浓度，该框架在基于细胞的模型中得到验证，并应用于肌强直性营养不良患者的肌肉。利用这一框架，我们评估了剪接事件的个体和组合预测人类活组织中功能性MBNL浓度的能力，以及它们作为生物标志物检测轻度、中度和重度糖尿病病例的性能。
Alternative splicing is a regulated process that results in expression of specific mRNA and protein isoforms. Alternative splicing factors determine the relative abundance of each isoform. Here we focus on MBNL1, a splicing factor misregulated in the disease myotonic dystrophy. By altering the concentration of MBNL1 in cells across a broad dynamic range, we show that different splicing events require different amounts of MBNL1 for half-maximal response, and respond more or less steeply to MBNL1. Motifs around MBNL1 exon 5 were studied to assess how cis-elements mediate the MBNL1 dose-dependent splicing response. A framework was developed to estimate MBNL concentration using splicing responses alone, validated in the cell-based model, and applied to myotonic dystrophy patient muscle. Using this framework, we evaluated the ability of individual and combinations of splicing events to predict functional MBNL concentration in human biopsies, as well as their performance as biomarkers to assay mild, moderate, and severe cases of DM.