Nonsense-mediated mRNA decay process in nine alleles of Niemann-Pick type C patients from Spain
Nonsense-mediated mRNA decay process in nine alleles of Niemann-Pick type C patients from Spain
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DOI：
10.1016/j.ymgme.2009.01.007
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发表时间：
2009-05-01
影响因子：
3.8
通讯作者：
Coll, Maria Josep
中科院分区：
文献类型：
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作者：
Macias-Vidal, Judit;Gort, Laura;Coll, Maria Josep
NPC1或NPC2基因突变是Niemann-Pick C型病（omim# 257220）的原因，这是一种常染色体隐性神经退行性溶酶体储存疾病，由细胞内脂质运输不调节引起。诸如无义或帧移位突变之类的改变会产生过早终止密码子（PTC）。无义介导的mRNA衰变（NMD）是一种自然的细胞过程，它降解编码过早截断蛋白的mRNA。在这项研究中，我们分析了产生PTC的9个NPC1突变（p.R116X, p.p q119vfsx8, pA&r260X, p.S425X, p.A558GfsXI2, p.p q775x, p.G993EfsX4, p.R1059X和p.p 11061nfsx4），以确定它们的mrna是否遭受NMD过程。为了实现这一目标，我们使用常规PCR和实时PCR比较了携带这些等位基因的患者在接受和未接受环己亚胺（CHX）治疗时的成纤维细胞。未经处理的成纤维细胞的常规PCR结果显示，与对照组相比，所有患者的NPC1 mRNA的量都有所减少。chx处理后，检测到mRNA的恢复，但不是在所有等位基因。然而，当使用实时PCR时，可以观察到包括那些定性显示没有明显增加的mRNA水平的等位基因的恢复。总之，我们证实NMD过程对所有分析的NPC1 ptc编码突变的mRNA衰减负责。(C) 2009爱思唯尔公司版权所有。
Mutations in NPC1 or NPC2 genes are responsible of Niemann-Pick type C disease (OMIM #257220), an autosomal recessive neurodegenerative lysosomal storage disorder caused by a non-regulation of intracellular lipid trafficking.Alterations such as nonsense or frame shift mutations generate a premature termination-codon (PTC). Nonsense-mediated mRNA decay (NMD) is a natural cellular process that degrades mRNAs that encode a prematurely truncated protein.In this study we have analyzed 9 NPC1 mutations which generate a PTC (p.R116X, p.Q119VfsX8, pA&r260X, p.S425X, p.A558GfsXI2, p.Q775X, p.G993EfsX4, p.R1059X and p.11061NfsX4), in order to determine if their mRNAs suffer NMD process. To achieve this objective we compared fibroblasts of patients carrying these alleles with and without cycloheximide (CHX) treatment using conventional PCR and real-time PCR.The results of conventional PCR of untreated fibroblasts showed a reduction of the amount of NPC1 mRNA compared to control in all patients. After CHX-treatment, a recovery of mRNA was detected but not in all the alleles. However, when real-time PCR was used, the recovery was observed including those alleles that qualitatively showed no apparent increase in mRNA level. In conclusion, we confirmed that NMD process is responsible for the mRNA decay for all the analyzed NPC1 PTC-encoding mutations. (C) 2009 Elsevier Inc. All rights reserved.