Nonsense-mediated mRNA decay in the ADAMTS13 gene caused by a 29-nucleotide deletion
Nonsense-mediated mRNA decay in the ADAMTS13 gene caused by a 29-nucleotide deletion
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由 29 个核苷酸缺失引起的 ADAMTS13 基因中无义介导的 mRNA 衰减
DOI：
10.3324/haematol.13102
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发表时间：
2008
期刊：
影响因子：
10.1
通讯作者：
F. Peyvandi
中科院分区：
文献类型：
--
作者：
I. Garagiola;C. Valsecchi;Silvia Lavoretano;H. Oren;M. Bohm;F. Peyvandi
本研究表明，两例严重的ADAMTS13缺乏是由两种不同的基因缺陷在两个不同的水平上作用的关联引起的。在哺乳动物细胞中，一种被称为无义介导的mRNA衰变的调节机制可以降解含有过早终止密码子的mRNA。该机制是内含子依赖性的，作为一种质量控制机制来消除异常转录本并调节各种自然发生的转录本的水平。在这项研究中，我们探索了两个复合杂合兄弟姐妹中ADAMTS13缺陷的分子机制，这些兄弟姐妹中一个等位基因携带一个29个核苷酸的缺失突变（c.291_319delGGAGGACACAGAGCGCTATGTGCTCACCA），另一个等位基因携带一个位于第二个CUB结构域的单碱基(a)插入突变（c.4143_4144insA）。采用实时定量逆转录酶聚合酶链式反应来探讨29个核苷酸缺失所引入的过早终止密码子是否触发了无义介导的mRNA衰变。结果体外表达研究表明，缺失29 bp后插入的过早终止密码子可能通过无义mRNA衰变的调控机制导致ADAMTS13 mRNA水平降低。此外，免疫荧光研究发现，4143_4144insA突变导致分泌受损，导致内质网中突变蛋白的保留。综上所述，本研究报道了ADAMTS13基因的两种不同缺陷是如何在两个不同水平上起作用的，即由29 bp缺失突变引起的过早终止密码子介导的衰变机制导致稳态mRNA水平受损，以及4143_4144insA导致分泌途径的改变，从而导致ADAMTS13严重缺失。
This study demonstrates that two cases of severe ADAMTS13 deficiency are mechanistically caused by the association of two different gene defects acting at two different levels. Background In mammalian cells a regulatory mechanism, known as nonsense-mediated mRNA decay, degrades mRNA harboring premature termination codons. This mechanism is intron-dependent and functions as a quality control mechanism to eliminate abnormal transcripts and modulates the levels of a variety of naturally occurring transcripts. Design and Methods In this study, we explored the molecular mechanism of ADAMTS13 deficiency in two compound heterozygous siblings carrying a 29-nucleotide deletion mutation located in exon 3 (c.291_319delGGAGGACACAGAGCGCTATGTGCTCACCA) in one allele and a single base (A) insertion mutation (c.4143_4144insA) in the second CUB domain previously reported in the other allele. Real-time quantitative reverse transcriptase polymerase chain reaction was used to explore whether the premature termination codons introduced by the deletion of the 29 nucleotides triggered the nonsense-mediated mRNA decay. Results In vitro-expression studies demonstrated that the premature termination codons inserted by the 29 bp deletion probably lead to a reduction of ADAMTS13 mRNA levels through the regulatory mechanisms of nonsense-mRNA decay. Furthermore, the 4143_4144insA mutation causes an impairment of secretion that leads to retention of the mutant protein in the endoplasmic reticulum, as observed in immunofluorescence studies. Conclusions In conclusion, this work reports how two different ADAMTS13 gene defects acting at two different levels, i.e, impairment of steady-state mRNA level caused by the premature termination codon mediated decay mechanism induced by the 29 bp deletion mutation and alteration of the secretion pathway due to 4143_4144insA, lead to a severe deficiency of ADAMTS13.
DOI：
10.1073/pnas.87.16.6306
发表时间：
1990-08-01
影响因子：
11.1
作者：
DENT, JA;BERKOWITZ, SD;RUGGERI, ZM
通讯作者：
RUGGERI, ZM
影响因子：
158.5
作者：
Tsai, HM;Lian, ECY
通讯作者：
Lian, ECY