Detection of two tissue-specific DNA-binding proteins with affinity for sites in the mouse beta-globin intervening sequence 2.

Detection of two tissue-specific DNA-binding proteins with affinity for sites in the mouse beta-globin intervening sequence 2.
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检测与小鼠 β-珠蛋白间插序列 2 中的位点具有亲和力的两种组织特异性 DNA 结合蛋白。

DOI:
10.1128/mcb.8.1.381-392.1988
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发表时间:
1988
影响因子:
5.3
通讯作者:
Housman,DE
Housman,DE
中科院分区:
生物学2区
文献类型:
--
作者:
Galson,DL;Housman,DE

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为了鉴定来自未诱导的小鼠红白血病核提取物的蛋白质,其特异性结合来自小鼠β-珠蛋白间插序列2(IVS 2)内的DNA酶I-超敏区的序列,使用凝胶电泳迁移率变动分析。检测到两种不同的序列特异性结合蛋白。这些因子的特异性结合位点通过DNA酶I保护足迹法和甲基化干扰法来确定。因子B1特异性结合两个同源位点,B1-A和B1-B,在IVS 2内和相反链上相距约100个碱基对。这两个区域可以独立地与因子B1相互作用。因子B1仅限于造血谱系细胞。因子B2结合于距离B1-A位点约5个碱基对的位点,并且仅限于红系细胞。这些因子的有限组织分布及其结合位点的位置表明,这些因子中的一种或两种可能参与小鼠β-珠蛋白基因IVS 2中组织特异性DNA酶I超敏位点的形成。
To identify proteins from uninduced murine erythroleukemia nuclear extracts which specifically bind to sequences from the DNase I-hypersensitive region within the mouse β-globin intervening sequence 2 (IVS2), a gel electrophoretic mobility shift assay was used. Two distinct sequence-specific binding proteins were detected. The specific binding sites for these factors were delineated by both DNase I protection footprinting and methylation interference. Factor B1 bound specifically to two homologous sites, B1-A and B1-B, approximately 100 base pairs apart within the IVS2 and on opposite strands. These two regions could interact with factor B1 independently. Factor B1 was limited to cells of hematopoietic lineages. Factor B2 bound to a site approximately 5 base pairs away from the B1-A site and was limited to cells of the erythroid lineage. The limited tissue distribution of these factors and the locations of their binding sites suggest that one or both of these factors may be involved in the formation of the tissue-specific DNase I-hypersensitive site in the IVS2 of the mouse β-globin gene.
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