DIFFERENTIAL BINDING OF THE NFE3 AND CP1/NFY TRANSCRIPTION FACTORS TO THE HUMAN GAMMA-GLOBIN AND EPSILON-GLOBIN CCAAT BOXES
DIFFERENTIAL BINDING OF THE NFE3 AND CP1/NFY TRANSCRIPTION FACTORS TO THE HUMAN GAMMA-GLOBIN AND EPSILON-GLOBIN CCAAT BOXES
复制标题
DOI：
10.1074/jbc.270.37.21934
复制
发表时间：
1995-09-15
影响因子：
4.8
通讯作者：
SANTORO, C
中科院分区：
文献类型：
--
作者：
RONCHI, AE;BOTTARDI, S;SANTORO, C
自然发生的非缺失突变影响人类γ-珠蛋白基因启动子的远端CCAAT盒，导致胎儿血红蛋白在成年后遗传性持续存在。尽管远端 CCAAT 盒是多种因素的靶标，包括 CP1/NFY、CDP、GATA-1 和 NFE3，但只有 NFE3 结合活性始终对该区域中明确表征的突变敏感，例如 G(-117) --> A、C-114 --> T 和胎儿血红蛋白的 Delta 13 遗传持久性。我们广泛表征了 NFE3 的结合特异性，并证明 NFE3 相对于其他 CCAAT 盒结合蛋白具有独特的特性。来自红系 K562 细胞的亲和纯化 NFE3 结合远端但不结合近端人 γ 珠蛋白 CCAAT 盒、人 ε 珠蛋白启动子的单个 CCAAT 盒以及进化相关的 Galago crassicaudatus γ 珠蛋白基因的近端 CCAAT 盒。在ε-球蛋白CCAAT盒内，NFE3代表主要且几乎唯一的结合活性。这种结合位点的破坏基本上使ε-珠蛋白启动子失活，这表明NFE3在该基因的胚胎表达中发挥着重要作用。
Naturally occurring nondeletional mutations affecting the distal CCAAT box of the human gamma-globin gene promoter result in hereditary persistence of fetal hemoglobin in adult life. Although the distal CCAAT box is the target of several factors, including CP1/NFY, CDP, GATA-1 and NFE3, only NFE3 binding activity is consistently sensitive to well characterized mutations in this region such as G(-117) --> A, C-114 --> T, and Delta 13 hereditary persistence of fetal hemoglobin. We extensively characterized the binding specificities of NFE3 and demonstrated that NFE3 has unique properties with respect to other CCAAT box-binding proteins. Affinity-purified NFE3 from erythroid K562 cells binds the distal but not the proximal human gamma-globin CCAAT box, the single CCAAT box of the human epsilon-globin promoter, and the proximal CCAAT box of the evolutionarily related Galago crassicaudatus gamma-globin gene. Within the epsilon-globin CCAAT box, NFE3 represents the major and almost exclusive binding activity. Disruption of such a binding site essentially inactivates the epsilon-globin promoter, suggesting that NFE3 plays an important role in the embryonic expression of this gene.