遗传 ›› 2017, Vol. 39 ›› Issue (3): 232-240.doi: 10.16288/j.yczz.16-317

• 研究报告 • 上一篇    下一篇

人类珠蛋白相关基因上游开放阅读框的变异分析

叶宇华(),张倩倩,钟建美,黎倚红,张立,喻秋霞,徐湘民()   

  1. 南方医科大学,基础医学院医学遗传学教研室,广州 510515
  • 收稿日期:2016-09-14 修回日期:2016-11-26 出版日期:2017-03-20 发布日期:2017-01-12
  • 作者简介:叶宇华,博士研究生,专业方向:医学遗传学。E-mail: biojamie@163.com|徐湘民,教授,博士生导师,研究方向:医学遗传学。E-mail: gzxuxm@pub.guangzhou.gd.cn
  • 基金资助:
    教育部博士点基金优先资助领域重点项目(20134433130001);国家自然科学基金委员会–广东省人民政府联合基金课题(U1201222)

Analysis of variants in upstream open reading frames of
human globin-related genes

Yuhua Ye(),Qianqian Zhang,Jianmei Zhong,Yihong Li,Li Zhang,Qiuxia Yu,Xiangmin Xu()   

  1. Department of Medical Genetics, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China
  • Received:2016-09-14 Revised:2016-11-26 Online:2017-03-20 Published:2017-01-12
  • Supported by:
    the the Doctoral Fund of Ministry of Education of China-Key Program of Priority Fields(20134433130001);the National Natural Science Foundation of China-Guangdong Joint Fund(U1201222)

摘要:

β-地中海贫血症是一种珠蛋白生成障碍的常染色体隐性遗传病。而γ珠蛋白基因的开放表达和胎儿血红蛋白的合成,是缓解β地中海贫血病人临床表型的一个重要因素。本研究针对202个血红蛋白相关调控基因或miRNA,对1802个β-地中海贫血症患者进行了目标区域捕获测序。通过生物信息学的分析,检测出了所有捕获区域内的突变。进一步对位于5′端非编码区(5′untranslated region, 5′UTR)的突变进行系统扫描,共计寻找到41个影响uORF(upstream open reading frame, uORF)有或无的功能性突变。从中选取了CHTOP基因(chr1:153606541 C>T)和TGFB1基因(chr19:41859418 G>A)的两个突变,通过定点诱变和双荧光素酶报告实验,在体外证实这两个突变均可显著地改变下游基因的表达。该研究结果为β-地中海贫血症临床表型的精确诊断提供了潜在的筛查靶点。

关键词: β-地中海贫血, 突变, 5′端非编码区, 上游开放阅读框

Abstract:

β-thalassemia is an autosomal recessive monogenic disease that is caused by defects in the production of β-like globin chains. Activation of γ-globin gene and the increase in fetal hemoglobin expression have been demonstrated as one of the most important factors to ameliorate the clinical outcome of β-thalassemia patients. In this study, 202 genes or miRNAs associated with human hemoglobin gene expression from 1802 β-thalassemia patients were analyzed with target capture and next generation sequencing strategies in terms of functional variants that might affect hemoglobin gene expression. The subsequent bioinformatics analysis included assessments of sequence quality, the variants within the target regions and the 5′UTR with potential effects on upstream open reading frames (uORFs). Among the 41 variants in 5′UTR potentially affecting the uORFs identified in the study, two variants (chr19: 41859418 G>A and chr1:153606541 C>T) were experimentally validated with dual-luciferase assays to be capable of significantly down-regulating the expression of TGFB1 and CHTOP gene, respectively. The present study demonstrated a system suitable for evaluating the importance of variants in 5′UTRs affecting uORFs in 202 human genes associated with hemoglobin expression. Research with this approach could provide potential targets that may contribute to the clinical phenotypes and provide biomarkers for precise diagnosis of β-thalassemia.

Key words: β-thalassemia, mutation, 5′untranslated region, upstream open reading frame