Multiple transcripts of the CYP21 gene are generated by the mutation of the splicing donor site in nitron 2 from GT to AT in 21-hydroxylase deficiency

H. H. Lee, S. F. Chang

研究成果: 雜誌貢獻文章

12 引文 (Scopus)

摘要

Maturation of primary RNA transcripts of eukaryotic genes often involves the removal of introns and joining of exons. The fidelity of RNA splicing is dependent on the identity of the nucleotide (nt) sequences at exon/intron boundaries. Most importantly, the highly conserved intronic 5′GT and 3′AG sequences are essential for correct splicing. Substitution of GT by any other nt leads to incomplete mRNA and a disruption of protein structure. We describe here the results of our transfection experiments in COS-1 cells with a CYP21 genomic construct that contained an IVS 2 + 1G→A mutation. Analysis of the transcripts by RT-PCR revealed that two different transcripts were generated by this mutant genome. In all the splicing products, we found that the entire exon 2 was deleted. Surprisingly, 30% of the transcripts from this mutant CYP21 genome were accompanied by an inclusion of 3′ intron 2 sequences due to the use of a different splice acceptor site. This is the first report of the molecular characterization of a splice donor site mutation in CYP21 via transcription in COS-1 cells.

原文英語
頁(從 - 到)397-402
頁數6
期刊Journal of Endocrinology
171
發行號3
DOIs
出版狀態已發佈 - 2001
對外發佈Yes

指紋

Introns
Exons
RNA Splice Sites
COS Cells
Mutation
Genome
Genes
RNA Splicing
RNA Precursors
Transfection
Nucleotides
Polymerase Chain Reaction
Messenger RNA
rice nuclear protein GT-2
Congenital adrenal hyperplasia due to 21 hydroxylase deficiency
Proteins

ASJC Scopus subject areas

  • Endocrinology

引用此文

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title = "Multiple transcripts of the CYP21 gene are generated by the mutation of the splicing donor site in nitron 2 from GT to AT in 21-hydroxylase deficiency",
abstract = "Maturation of primary RNA transcripts of eukaryotic genes often involves the removal of introns and joining of exons. The fidelity of RNA splicing is dependent on the identity of the nucleotide (nt) sequences at exon/intron boundaries. Most importantly, the highly conserved intronic 5′GT and 3′AG sequences are essential for correct splicing. Substitution of GT by any other nt leads to incomplete mRNA and a disruption of protein structure. We describe here the results of our transfection experiments in COS-1 cells with a CYP21 genomic construct that contained an IVS 2 + 1G→A mutation. Analysis of the transcripts by RT-PCR revealed that two different transcripts were generated by this mutant genome. In all the splicing products, we found that the entire exon 2 was deleted. Surprisingly, 30{\%} of the transcripts from this mutant CYP21 genome were accompanied by an inclusion of 3′ intron 2 sequences due to the use of a different splice acceptor site. This is the first report of the molecular characterization of a splice donor site mutation in CYP21 via transcription in COS-1 cells.",
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N2 - Maturation of primary RNA transcripts of eukaryotic genes often involves the removal of introns and joining of exons. The fidelity of RNA splicing is dependent on the identity of the nucleotide (nt) sequences at exon/intron boundaries. Most importantly, the highly conserved intronic 5′GT and 3′AG sequences are essential for correct splicing. Substitution of GT by any other nt leads to incomplete mRNA and a disruption of protein structure. We describe here the results of our transfection experiments in COS-1 cells with a CYP21 genomic construct that contained an IVS 2 + 1G→A mutation. Analysis of the transcripts by RT-PCR revealed that two different transcripts were generated by this mutant genome. In all the splicing products, we found that the entire exon 2 was deleted. Surprisingly, 30% of the transcripts from this mutant CYP21 genome were accompanied by an inclusion of 3′ intron 2 sequences due to the use of a different splice acceptor site. This is the first report of the molecular characterization of a splice donor site mutation in CYP21 via transcription in COS-1 cells.

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