Effect of splice-site polymorphisms of theTMPRSS4, NPHP4 andORCTL4 genes on their mRNA expression
Effect of splice-site polymorphisms of theTMPRSS4, NPHP4 andORCTL4 genes on their mRNA expression
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DOI：
10.1007/bf02715838
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发表时间：
2005-08
影响因子：
1.5
通讯作者：
Hidetaka Yamada;K. Shinmura;T. Tsuneyoshi;H. Sugimura
中科院分区：
文献类型：
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作者：
Hidetaka Yamada;K. Shinmura;T. Tsuneyoshi;H. Sugimura
与其基因产物的结构变化相关的遗传多态性就其与疾病的潜在关系而言是重要的。因此，在这项研究中，跨膜丝氨酸蛋白酶基因TMPRSS 4，肾单位结核基因NPHP 4，和有机阳离子转运蛋白基因ORCTL 4的剪接位点变异体，从dbSNP单核苷酸多态性数据库中选择的候选人，以确定与其mRNA转录的结构变化相关的遗传多态性。用PCR-SSCP方法分析日本人群TMPRSS4c.4 -7A> G、NPHP4c.2818 -2A>T和ORCTL4c.517 -2A>C多态性的等位基因频率分别为0.42、0.10和0.27。接下来，通过RT-PCR分析和测序分析研究这些多态性对前体mRNA剪接模式的影响。TMPRSS 4、NPHP 4和ORCTL 4多态性分别与r.4-6_4- 1 ins转录本、r.2818_2823del和r.2818_2859del转录本以及r.517-94_517- 1 ins; r.517-2a>c和r.517_620del转录本的产生相关。由于所有这些转录物编码的蛋白质都与氨基酸插入/缺失和过早终止形式的相对显著的结构变化相关，因此它们的功能能力可能会大大降低。我们证明，由于剪接位点多态性而导致mRNA转录物的结构变化，这意味着它们在某些病理条件下可能具有生物学意义。
Genetic polymorphisms associated with structural changes of their gene product are important in terms of their potential relation with diseases. Therefore, in this study, splice-site variants of the transmembrane serine protease geneTMPRSS4, nephronophthisis geneNPHP4, and organic-cation transporter geneORCTL4, were selected from the dbSNP single nucleotide polymorphism database as candidates to identify genetic polymorphisms associated with a structural change in their mRNA transcripts. The allele frequencies of theTMPRSS4c.4-7A>G,NPHP4c.2818-2A>T, andORCTL4c.517-2A>C polymorphisms in a Japanese population were determined to be 0.42, 0.10, and 0.27, respectively, by PCR-SSCP analysis. Next, the effect of these polymorphisms on the mode of pre-mRNA splicing was investigated by RT-PCR analysis followed by sequencing analysis. TheTMPRSS4, NPHP4, andORCTL4polymorphisms were associated with the production of the r.4-6_4-1ins transcript, the r.2818_2823del and r.2818_2859del transcripts, and the r.517-94_517-1ins; r.517-2a>c and r.517_620del transcripts, respectively. Since the proteins encoded by all these transcripts are associated with relatively significant structural changes in the form amino acid insertion/deletion and premature termination, their functional ability may be greatly reduced. Our demonstration of structural changes in mRNA transcripts as a result of splice-site polymorphisms implies that they may be of biological significance in certain pathological conditions.