Mutation of a barrier insulator in the human ankyrin-1 gene is associated with hereditary spherocytosis
Mutation of a barrier insulator in the human ankyrin-1 gene is associated with hereditary spherocytosis
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DOI：
10.1172/jci42240
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发表时间：
2010-12-01
影响因子：
15.9
通讯作者：
Bodine, David M.
中科院分区：
文献类型：
--
作者：
Gallagher, Patrick G.;Steiner, Laurie A.;Bodine, David M.
锚蛋白-1基因缺陷是人类遗传性球形红细胞增多症最常见的原因，这是一种影响所有种族患者的遗传性贫血。在某些激酶中，在红系细胞中有活性的锚蛋白基因启动子的上游区域发现了连锁的-108/-153核苷酸取代。在体内，锚蛋白红细胞启动子及其上游区域指导位置独立的、均匀的表达，具有屏障绝缘体的性质。使用人红系细胞系和原代细胞和转基因小鼠，在这里，我们已经证明，红系启动子的上游区域是体内红系细胞中的屏障绝缘体。该区域表现出屏障的功能和结构特征，包括在体内功能测定中防止基因沉默、适当的染色质构型和被屏障相关蛋白占据。具有-108/-153球形红细胞增多相关突变的片段在体内不能作为屏障绝缘体，并表现出屏障相关染色质构型的扰动。在转基因小鼠中，侧翼突变的-108/-153锚蛋白基因启动子与充分表征的鸡HS 4屏障绝缘子恢复了与野生型相当的水平的位置无关的均匀表达。这些数据表明，锚蛋白-1类红细胞启动子的上游区域作为一个屏障绝缘体，并确定破坏的屏障元件作为一个潜在的人类疾病的发病机制。
Defects of the ankyrin-1 gene are the most common cause in humans of hereditary spherocytosis, an inherited anemia that affects patients of all ethnic groups. In some kindreds, linked -108/-153 nucleotide substitutions have been found in the upstream region of the ankyrin gene promoter that is active in erythroid cells. In vivo, the ankyrin erythroid promoter and its upstream region direct position-independent, uniform expression, a property of barrier insulators. Using human erythroid cell lines and primary cells and transgenic mice, here we have demonstrated that a region upstream of the erythroid promoter is a barrier insulator in vivo in erythroid cells. The region exhibited both functional and structural characteristics of a barrier, including prevention of gene silencing in an in vivo functional assay, appropriate chromatin configuration, and occupancy by barrier- associated proteins. Fragments with the -108/-153 spherocytosis-associated mutations failed to function as barrier insulators in vivo and demonstrated perturbations in barrier-associated chromatin configuration. In transgenic mice, flanking a mutant -108/-153 ankyrin gene promoter with the well-characterized chicken HS4 barrier insulator restored position-independent, uniform expression at levels comparable to wild-type. These data indicate that an upstream region of the ankyrin-1 erythroid promoter acts as a barrier insulator and identify disruption of the barrier element as a potential pathogenetic mechanism of human disease.