Chu C S, Trapnell B C, Murtagh J J, Moss J, Dalemans W, Jallat S, Mercenier A, Pavirani A, Lecocq J P, Cutting G R
Pulmonary Branch, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD 20892.
EMBO J. 1991 Jun;10(6):1355-63. doi: 10.1002/j.1460-2075.1991.tb07655.x.
The predicted protein domains coded by exons 9-12 and 19-23 of the 27 exon cystic fibrosis transmembrane conductance regulator (CFTR) gene contain two putative nucleotide-binding fold regions. Analysis of CFTR mRNA transcripts in freshly isolated bronchial epithelium from 12 normal adult individuals demonstrated that all had some CFTR mRNA transcripts with exon 9 completely deleted (exon 9- mRNA transcripts). In most (9 of 12), the exon 9- transcripts represented less than or equal to 25% of the total CFTR transcripts. However, in three individuals, the exon 9- transcripts were more abundant, comprising 39, 62 and 66% of all CFTR transcripts. Re-evaluation of the same individuals 2-4 months later showed the same proportions of exon 9- transcripts. Of the 24 CFTR alleles in the 12 individuals, the sequences of the exon-intron junctions relevant to exon 9 deletion (exon 8-intron 8, intron 8-exon 9, exon 9-intron 9, and intron 9-exon 10) were identical except for the intron 8-exon 9 region sequences. Several individuals had varying lengths of a TG repeat in the region between splice branch and splice acceptor consensus sites. Interestingly, one allele in each of the two individuals with 62 and 66% exon 9- transcripts had a TT deletion in the splice acceptor site for exon 9. These observations suggest either the unlikely possibility that sequences in exon 9 are not critical for the functioning of the CFTR or that only a minority of the CFTR mRNA transcripts need to contain exon 9 sequences to produce sufficient amounts of a normal CFTR to maintain a normal clinical phenotype.
由27个外显子组成的囊性纤维化跨膜传导调节因子(CFTR)基因的外显子9 - 12和19 - 23编码的预测蛋白质结构域包含两个假定的核苷酸结合折叠区域。对12名正常成年个体新鲜分离的支气管上皮中的CFTR mRNA转录本进行分析,结果显示所有人都有一些CFTR mRNA转录本,其中外显子9完全缺失(外显子9 - mRNA转录本)。在大多数个体(12名中的9名)中,外显子9 - 转录本占CFTR转录本总数的比例小于或等于25%。然而,在三名个体中,外显子9 - 转录本更为丰富,分别占所有CFTR转录本的39%、62%和66%。在2 - 4个月后对相同个体进行重新评估,结果显示外显子9 - 转录本的比例相同。在这12名个体的24个CFTR等位基因中,与外显子9缺失相关的外显子 - 内含子连接序列(外显子8 - 内含子8、内含子8 - 外显子9、外显子9 - 内含子9和内含子9 - 外显子10)除内含子8 - 外显子9区域序列外均相同。几名个体在剪接分支和剪接受体共有序列之间的区域有不同长度的TG重复序列。有趣的是,在外显子9 - 转录本占62%和66%的两名个体中,各自有一个等位基因在外显子9的剪接受体位点存在TT缺失。这些观察结果表明,要么外显子9中的序列对CFTR的功能并非至关重要这种可能性不大,要么只有少数CFTR mRNA转录本需要包含外显子9序列才能产生足够量的正常CFTR以维持正常的临床表型。