Liebhaber S A, Cash F E, Main D M
J Clin Invest. 1985 Sep;76(3):1057-64. doi: 10.1172/JCI112058.
alpha-Globin is encoded by the two adjacent genes, alpha 1 and alpha 2. Although it is clearly established that both alpha-globin genes are expressed, their relative contributions to alpha-globin messenger RNA (mRNA) and protein synthesis are not fully defined. Furthermore, changes that may occur in alpha-globin gene activity secondarily to the loss of function of one or more of these genes (alpha-thalassemia [Thal]) have not been directly investigated. This study further defines the expression of the two human alpha-globin genes by determining the relative levels of alpha 1 and alpha 2 mRNA in the reticulocytes of normal individuals and in individuals heterozygous for the common 3.7-kilobase deletion within the alpha-globin gene cluster that removes the alpha 2-globin gene (the rightward type alpha-Thal-2 deletion). To quantitate accurately the ratio of the two alpha-globin mRNAs, we have modified a previously reported S1 nuclease assay to include the use of 32P end-labeled probes isolated from alpha 1- and alpha 2-globin complementary DNA recombinant plasmids. In individuals with a normal alpha-globin genotype (as determined by Southern blot analysis [alpha alpha/alpha alpha]), alpha 2-globin mRNA is present at an average 2.8-fold excess to alpha 1. In individuals heterozygous for the rightward type alpha-Thal-2 deletion (-alpha/alpha alpha) the alpha 2/alpha 1 mRNA ratio is 1:1. These results suggest that the loss of the alpha 2-globin gene in the alpha-Thal-2 deletion is associated with a 1.8-fold compensatory increase alpha 1-globin gene expression.
α-珠蛋白由两个相邻基因α1和α2编码。尽管已明确两个α-珠蛋白基因均有表达,但其对α-珠蛋白信使核糖核酸(mRNA)和蛋白质合成的相对贡献尚未完全明确。此外,这些基因中的一个或多个功能丧失(α-地中海贫血[Thal])继发的α-珠蛋白基因活性变化尚未得到直接研究。本研究通过测定正常个体以及α-珠蛋白基因簇内常见3.7千碱基缺失(该缺失去除了α2-珠蛋白基因,即向右型α-Thal-2缺失)的杂合个体网织红细胞中α1和α2 mRNA的相对水平,进一步明确了两个人类α-珠蛋白基因的表达情况。为了准确量化两种α-珠蛋白mRNA的比例,我们改进了先前报道的S1核酸酶测定法,使其包括使用从α1-和α2-珠蛋白互补DNA重组质粒中分离的32P末端标记探针。在α-珠蛋白基因型正常的个体(通过Southern印迹分析确定为[αα/αα])中,α2-珠蛋白mRNA的平均含量比α1多2.8倍。在向右型α-Thal-2缺失的杂合个体(-α/αα)中,α2/α1 mRNA比例为1:1。这些结果表明,α-Thal-2缺失中α2-珠蛋白基因的缺失与α1-珠蛋白基因表达的1.8倍代偿性增加有关。