Dyer M A, Naidoo R, Hayes R J, Larson C J, Verdine G L, Baron M H
Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts 02138, USA.
Mol Cell Biol. 1996 Mar;16(3):829-38. doi: 10.1128/MCB.16.3.829.
The mammalian beta-like globin gene family has served as an important model system for analysis of tissue- and developmental state-specific gene regulation. Although the activities of a number of regulatory proteins have been implicated in the erythroid cell-specific transcription of globin genes, the mechanisms that restrict their expression to discrete stages of development are less well understood. We have previously identified a novel regulatory element (PRE II) upstream from the human embryonic beta-like globin gene (epsilon) that synergizes with other sequences to confer tissue- and stage-specific expression on a minimal epsilon-globin gene promoter in cultured embryonic erythroid cells. Binding of an erythroid nuclear protein (PRE II-binding factor [PRE-IIBF]) to the PRE II control element is required for promoter activation. Here we report on some of the biochemical properties of PREIIBF, including the characterization of its specificity and affinity for DNA. The embryonic and adult forms of PREIIBF recognize their cognate sequences with identical specificities, supporting our earlier conclusion that they are very similar proteins. PREIIBF binds DNA as a single polypeptide with an Mr of approximately 80,000 to 85,000 and introduces a bend into the target DNA molecule. These results suggest a mechanism by which PREIIBF may contribute to the regulation of the embryonic beta-like globin gene within the context of a complex locus.
哺乳动物β样珠蛋白基因家族一直是分析组织和发育状态特异性基因调控的重要模型系统。尽管许多调控蛋白的活性与珠蛋白基因的红系细胞特异性转录有关,但将它们的表达限制在离散发育阶段的机制却知之甚少。我们之前在人类胚胎β样珠蛋白基因(ε)上游鉴定出一个新型调控元件(PRE II),它与其他序列协同作用,使培养的胚胎红系细胞中的最小ε珠蛋白基因启动子具有组织和阶段特异性表达。启动子激活需要一种红系核蛋白(PRE II结合因子[PRE-IIBF])与PRE II调控元件结合。在此,我们报道了PREIIBF的一些生化特性,包括其对DNA的特异性和亲和力的表征。PREIIBF的胚胎型和成年型以相同的特异性识别其同源序列,支持了我们之前的结论,即它们是非常相似的蛋白质。PREIIBF作为一种分子量约为80,000至85,000的单一多肽结合DNA,并使靶DNA分子产生弯曲。这些结果提示了一种机制,通过该机制PREIIBF可能在复杂基因座的背景下参与胚胎β样珠蛋白基因的调控。