DIFFERENTIAL SPLICING OF COL4A5 MESSENGER-RNA IN KIDNEY AND WHITE BLOOD-CELLS - A COMPLEX MUTATION IN THE COL4A5 GENE OF AN ALPORT PATIENT DELETES THE NC1 DOMAIN
DIFFERENTIAL SPLICING OF COL4A5 MESSENGER-RNA IN KIDNEY AND WHITE BLOOD-CELLS - A COMPLEX MUTATION IN THE COL4A5 GENE OF AN ALPORT PATIENT DELETES THE NC1 DOMAIN
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DOI：
10.1038/ki.1993.384
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发表时间：
1993-12-01
影响因子：
19.6
通讯作者：
MARYNEN, P
中科院分区：
文献类型：
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作者：
GUO, CY;VANDAMME, B;MARYNEN, P
优化PCR条件以从成淋巴细胞和肾组织中扩增COL 4A 5 cDNA。从Alport综合征患者的肾脏中分离的COL 4A 5 mRNA的测序显示与公布的序列有两个差异。一个分歧是，在COL 4A 5 mRNA的外显子11和10之间插入18 bp序列，为COL 4A 5序列增加了两个Gly-X-Y三联体，随后在四个正常肾脏mRNA样品的mRNA中发现。该序列在我们测序的所有白色血细胞RNA样品中均不存在，表明组织特异性剪接，在肾脏COL 4A 5 mRNA中存在额外的外显子。肾脏和白色血细胞中COL 4A 5 mRNA差异剪接的这一发现可能会影响使用白色血细胞mRNA分析Alport突变。第二，在AS患者的mRNA中检测到复杂的突变，在信使中引入提前终止密码子，缺失部分三螺旋结构域和完整的NC结构域。患者的母亲被证明是这种突变的杂合子。
PCR conditions were optimized to amplify the COL4A5 cDNA from lymphoblasts and kidney tissue. Sequencing of the COL4A5 mRNA isolated from the kidney of an Alport syndrome patient revealed two differences with the published sequence. One divergence, the insertion of an 18 bp sequence between exon 11 and 10 of the COL4A5 mRNA added two Gly-X-Y triplets to the COL4A5 sequence and was subsequently found in the mRNA of four normal kidney mRNA samples. This sequence was absent in all white blood cell RNA samples sequenced by us, indicating tissue specific splicing with the presence of an additional exon in kidney COL4A5 mRNA. This finding of differential splicing of COL4A5 mRNA in kidney and white blood cells might affect the use of white blood cell mRNA for the analysis of Alport mutations. Second, a complex mutation was detected in the mRNA from the AS patient introducing a premature stop codon in the message, deleting part of the triple helical domain and the complete NC domain. The mother of the patient was shown to be heterozygous for this mutation.