Germinal HPRT splice donor site mutation results in multiple RNA splicing products in T-lymphocyte cultures
Germinal HPRT splice donor site mutation results in multiple RNA splicing products in T-lymphocyte cultures
复制标题
DOI：
10.1007/bf02369904
复制
发表时间：
1996-03-01
期刊：
影响因子：
--
通讯作者：
ONeill, JP
中科院分区：
文献类型：
--
作者：
Hunter, TC;Melancon, SB;ONeill, JP
我们利用外周血 T 淋巴细胞培养物分析了两名 Lesch-Nylan 综合征男性表兄弟的 hprt 突变，并确认了家庭中女性成员的携带者状态。 cDNA 和基因组 DNA 测序研究均表明，该患者在外显子 5 供体剪接位点序列中携带迄今未描述的单碱基缺失（15：+/- 1，Delta G，碱基号 31635），最大的 cDNA 产物包含所有 9 个 hprt 外显子以及内含子 5 的 66 个碱基插入，与内含子 5 中隐秘剪接位点的使用一致(aag(67)/gtaagc)。这种剪接错误将导致紧接着外显子 5 (15:2-4, taa) 后出现链终止密码子，并预测为 133 个氨基酸的多肽。正常剪接供体位点的这种丢失也会导致多种 hprt mRNA 种类，结合使用内含子 5 中的隐性剪接位点和涉及外显子 2-6 的剪接错误。除了定义新的 Lesch-Nylan 突变 (hpTt(Henryville)) 之外，这些结果还提供了对 T 淋巴细胞中 hprt mRNA 异常剪接的深入了解。
We have used peripheral blood T-lymphocyte cultures to analyze the hprt mutation in two Lesch-Nylan syndrome males who are cousins and to confirm the carrier status of female members of the family. Both cDNA and genomic DNA sequencing studies show that this patient carries a hitherto undescribed single base deletion in the exon 5 donor splice site sequence (15: +/- 1, Delta G, base number 31635), The largest cDNA product contained all nine hprt exons plus an insertion of 66 bases of intron 5, consistent with the use of a cryptic splice site in intron 5 (aag(67)/gtaagc). This splicing error would result in a chain terminating codon immediately after exon 5 (15:2-4, taa) and predicts a polypeptide of 133 amino acids, This loss of the normal splice donor site also results in multiple hprt mRNA species, combining the use of the cryptic splice site in intron 5 and splicing errors involving exons 2-6. In addition to defining a new Lesch-Nylan mutation (hpTt(Henryville)), these results provide insight into aberrant splicing of hprt mRNA in T-lymphocytes.