Three Common Polymorphisms in the CYBA Gene Form a Haplotype Associated With Decreased ROS Generation
Three Common Polymorphisms in the CYBA Gene Form a Haplotype Associated With Decreased ROS Generation
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DOI：
10.1002/humu.21029
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发表时间：
2009-07-01
期刊：
影响因子：
3.9
通讯作者：
Krause, Karl-Heinz
中科院分区：
文献类型：
--
作者：
Bedard, Karen;Attar, Homa;Krause, Karl-Heinz
NOX酶是产生活性氧（ROS）的NADPH氧化酶。NOX家族的几个成员依赖于CYBA基因编码的p22（phox）亚基。CYBA具有高度多态性，已被广泛研究为各种疾病的潜在危险因素，但结果相互矛盾。在本研究中，我们使用EB病毒（EBV）转化的B淋巴细胞从50个健康无关的个体，分析他们的CYBA mRNA序列和NOX 2依赖的ROS的产生。七个单核苷酸多态性（SNPs）被确定（五个以前描述过，两个新的）。这些SNPs的组合产生了11种不同的单倍型，可分为7个单倍群（A-G）。单倍群C（c.214T > C，c.521T > C，和c.521T > C）24 G> A）与最常见的单倍型群A相比，显示出显著较低的ROS产生。将来自七个单倍型群的CYBA变体转导到p22（phox）缺陷型B淋巴细胞中。单倍群C变异体表现出显著较低的ROS产生。c.214T > C和c.521T > C导致非同义密码子改变，而c.* 24 G> A位于3 'UTR内。使用荧光素酶/3 'UTR构建体，我们表明 * 24 A等位基因导致报告基因活性降低。这些结果有助于揭示CYBA遗传变异如何影响NOX 2活性的复杂本质，并表明单倍型而不是单个SNP定义了对ROS生成的影响。Mutat 30：1123-1133，2009. (C)2009威利-利斯公司
NOX enzymes are reactive oxygen species (ROS)-generating NADPH oxidases. Several members of the NOX family depend on the p22(phox) subunit, encoded by the CYBA gene. CYBA is highly polymorphic, and has been widely studied as a potential risk factor for various diseases, with conflicting results. In the present study, we used Epstein-Barr (EBV)-transformed B-lymphocytes from 50 healthy unrelated individuals to analyze their CYBA mRNA sequence and NOX2-dependent ROS generation. Seven single nucleotide polymorphisms (SNPs) were identified (five previously described, two novel). The combination of these SNPs yielded 11 distinct haplotypes, which could be grouped into seven haplogroups (A-G). Haplogroup C (c.214T > C, c.521T > C, and c.*24G > A) showed a significantly lower ROS generation, as compared to the most frequent haplogroup, A. CYBA variants from the seven haplogroups were transduced into p22(phox)-deficient B-lymphocytes. The haplogroup C variant showed significantly lower ROS production. c.214T > C and c.521T > C lead to nonsynonymous codon changes while c.*24G > A lies within the 3'UTR. Using a luciferase/3'UTR construct, we showed that the *24A allele led to decreased reporter gene activity. These results help to unravel the complex nature of how genetic variations in CYBA influence NOX2 activity, and indicate that haplotypes, rather than individual SNPs, define the effect on ROS generation. Hum Mutat 30:1123-1133, 2009. (C) 2009 Wiley-Liss, Inc.