Mutation screening at the RNA level of the STK11/LKB-1 gene in Peutz-Jeghers syndrome reveals complex splicing abnormalities and a novel mRNA isoform (STK11 c.597 ∧ 598insIVS4)
Mutation screening at the RNA level of the STK11/LKB-1 gene in Peutz-Jeghers syndrome reveals complex splicing abnormalities and a novel mRNA isoform (STK11 c.597 ∧ 598insIVS4)
复制标题
DOI：
10.1002/humu.1211
复制
发表时间：
2001-01-01
期刊：
影响因子：
3.9
通讯作者：
Ballhausen, WG
中科院分区：
文献类型：
--
作者：
Abed, AA;Günther, K;Ballhausen, WG
本研究旨在评价一种基于逆转录聚合酶链反应的蛋白质截短试验，用于鉴定Peutz-Jeghers综合征（PJS）丝氨酸/苏氨酸蛋白激酶11（STK 11，也称为LKB 1）基因的种系突变。我们的数据证实，STK 11的失活可能是由于不寻常的干扰剪接调节，导致截短的蛋白质。然而，无义介导的mRNA衰变必须用嘌呤霉素阻断，以检测PJS患者白细胞mRNA库中所含的缩短的STK 11基因产物。有趣的是，两个突变逃脱了检测外显子测序技术与通常的侧翼PCR引物，因为改变位于内含子序列的中间。我们描述了一个复合杂合子PJS患者携带两个不同的突变内含子1上的不同等位基因。这两种突变分别传给了他的两个孩子中的一个。在我们基于RNA的分析过程中，我们在各种组织中检测到保留内含子4的新型STK 11/LKB 1 mRNA变体（STK 11 c.597（boolean AND）598 insIVS 4）的高水平表达。这种mRNA亚型起始于另一个转录调控区，如引物延伸分析所示，即使在正常启动子完全甲基化的细胞系中也是如此。由于新的突变机制确定，我们讨论的影响，基于RNA的策略，检测在PJS的germinostK 11突变。2001年，《突变》18：397-410。(C)2001 Wiley-Liss，Inc.
This study was intended to evaluate a diagnostic reverse transcriptase polymerase chain reaction based protein-truncation test for the identification of germline mutations in the serine/threonine protein kinase 11 (STK11, also designated LKB1) gene in Peutz-Jeghers syndrome (PJS). Our data exemplify that the inactivation of STK11 can be due to unusual disturbances in splicing regulation which result in truncations of the protein. However, nonsense mediated mRNA decay must be blocked with puromycin to detect shortened STK11 gene products contained in the leucocytic mRNA pool of PJS patients. Interestingly, two mutations escaped from detection by exon sequencing techniques with usual flanking PCR primers, since alterations were located right in the middle of intronic sequences. We describe a compound heterozygous PJS patient who carried two different mutations in intron 1 on separate alleles. Each of the two mutations was transmitted individually to one of his two children. In the course of our RNA based analyses we detected high level expression of a novel STK11/LKB1 mRNA variant retaining intron 4 (STK11 c.597(boolean AND)598insIVS4) in various tissues. This mRNA isoform was initiated from an alternative transcription regulatory region as revealed by primer extension analyses even in cell lines with complete methylation of the normal promoter. As a consequence of novel mutational mechanisms identified we discuss the impact of RNA based strategies for the detection of germinal STK11 mutations in PJS. Hum Mutat 18:397-410, 2001. (C) 2001 Wiley-Liss, Inc.