格雷夫斯病患者甲状腺过氧化物酶信使核糖核酸中存在可变剪接的证据。

Evidence for an alternate splicing in the thyroperoxidase messenger from patients with Graves' disease.

作者信息

Zanelli E, Henry M, Charvet B, Malthièry Y

机构信息

Laboratoire de Biochimie Médicale, U.38 INSERM, Faculté de Médecine, Marseille, France.

出版信息

Biochem Biophys Res Commun. 1990 Jul 31;170(2):735-41. doi: 10.1016/0006-291x(90)92152-p.

DOI:10.1016/0006-291x(90)92152-p

PMID:2383265

Abstract

An initial lambda gt 11 cDNA library constructed from a human Graves' patient thyroid was screened with an immunopurified rabbit anti-human thyroperoxidase (hTPO) polyclonal antibody. A 869 bp clone was obtained. It presents a 130 bp deletion as compared to the published sequence and a 77 bp insertion in the 3' non-coding region. Screening of a pUC cDNA library from another Graves' patient thyroid exhibited the same 130 bp deletion in two other cDNA clones. PCR analysis of mRNA transcripts confirmed the presence of the two messengers in two other Graves' thyroid tissues. In all the cases, this new spliced mRNA species represents between 40% and 50% of the total hTPO mRNAs. With respect to the structure of the hTPO gene, the present deletion suggests an alternate splicing of exon 16. The juxtaposition of exon 17 to exon 15 encoding the transmembrane domain leads to a shift in the reading frame. By the use of a different stop codon, the spliced mRNA generates a modified 56 - COOH terminal aminoacids (aa) sequence.

摘要

用人Graves病患者甲状腺构建初始λgt 11 cDNA文库，并用免疫纯化的兔抗人甲状腺过氧化物酶（hTPO）多克隆抗体进行筛选。获得一个869 bp的克隆。与已发表序列相比，它存在130 bp的缺失，且在3'非编码区有77 bp的插入。对另一位Graves病患者甲状腺的pUC cDNA文库进行筛选，在另外两个cDNA克隆中也显示出相同的130 bp缺失。对mRNA转录本进行PCR分析，证实另外两个Graves病甲状腺组织中存在这两种信使RNA。在所有情况下，这种新的剪接mRNA种类占hTPO总mRNA的40%至50%。关于hTPO基因的结构，目前的缺失提示外显子16的可变剪接。编码跨膜结构域的外显子17与外显子15并列导致阅读框移位。通过使用不同的终止密码子，剪接后的mRNA产生一个修饰的56 - COOH末端氨基酸（aa）序列。