Unraveling the molecular basis underlying nine putative splice site variants of von Willebrand factor
Unraveling the molecular basis underlying nine putative splice site variants of von Willebrand factor
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揭示冯维勒布兰德因子九个假定剪接位点变异的分子基础
DOI：
10.1002/humu.24312
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发表时间：
2021-12-17
期刊：
影响因子：
3.9
通讯作者：
Wang, Xuefeng
中科院分区：
文献类型：
--
作者：
Liang, Qian;Lin, Xiaoyi;Wang, Xuefeng
大约10%的von Willebrand因子(VWF)基因变异体被怀疑扰乱信使RNA(MRNA)的加工，由于缺乏转录分析，其数量可能被低估。在本研究中，我们提供了一种详细的策略来评估VWF的9个可能的剪接位点变体(PSSV)对mRNA加工和蛋白质性质的影响，并建立它们的基因-表型关系。9个PSSV中有8个影响VWF剪接：C.322A>；T、c.1534-13_1551delinsCA和c.8116-2del引起外显子跳跃；c.221-2A>；C、c.323+1G>；T和c.2547-13T>；A导致神秘剪接位点的激活；c.2684A>；G导致外显子跳跃并激活一个神秘剪接位点；c.2968-14A>；G创建了一个新的剪接位点。其余的c.5171-9del可能是良性的。与白细胞相比，无意义介导的mRNA衰变(NMD)在血小板中的效率要高得多，这损害了8个PSSV中4个异常转录本的识别。无义突变体c.322A>；T部分损害了mRNA的加工，泄漏了与c.322T(p.Arg108*)的少量正确转录本，而错义突变体c.2684A>；G完全扰乱了VWF的正常剪接，而不是产生了用Gln895Arg替代的突变蛋白。这项研究的结果肯定会增加对von Willebrand病背后的分子事件的新见解。
Approximately 10% of von Willebrand factor (VWF) gene variants are suspected to disrupt messenger RNA (mRNA) processing, the number of which might be underestimated due to the lack of transcript assays. In the present study, we provided a detailed strategy to evaluate the effects of nine putative splice site variants (PSSVs) of VWF on mRNA processing as well as protein properties and establish their genotype-phenotype relationships. Eight of nine PSSVs affected VWF splicing: c.322A>T, c.1534-13_1551delinsCA, and c.8116-2del caused exon skipping; c.221-2A>C, c.323+1G>T, and c.2547-13T>A resulted in the activation of cryptic splice sites; c.2684A>G led to exon skipping and activation of a cryptic splice site; c.2968-14A>G created a new splice site. The remaining c.5171-9del was likely benign. The efficiency of nonsense-mediated mRNA decay (NMD) was much higher in platelets compared to leukocytes, impairing the identification of aberrant transcripts in 4 of 8 PSSVs. The nonsense variant c.322A>T partially impaired mRNA processing, leaking a small amount of correct transcripts with c.322T (p.Arg108*), while the missense variant c.2684A>G totally disrupted normal splicing of VWF, rather than produced mutant protein with the substitution of Gln895Arg. The results of this study would certainly add novel insights into the molecular events behind von Willebrand disease.