Inducible DNA-protein interactions of the murine kappa immunoglobulin enhancer in intact cells: comparison with in vitro interactions.
Inducible DNA-protein interactions of the murine kappa immunoglobulin enhancer in intact cells: comparison with in vitro interactions.
复制标题
完整细胞中小鼠 kappa 免疫球蛋白增强剂的诱导 DNA-蛋白质相互作用：与体外相互作用的比较。
DOI：
10.1093/nar/16.3.953
复制
发表时间：
1988
影响因子：
14.9
通讯作者：
VanNess,B
中科院分区：
文献类型：
--
作者：
Hromas,R;Pauli,U;Marcuzzi,A;Lafrenz,D;Nick,H;Stein,J;Stein,G;VanNess,B
κ免疫球蛋白基因的大内含子含有一个顺式作用的增强子元件，它在基因的组织特异性表达中起重要作用。我们已经证实了从表达或诱导表达κ基因的细胞系中提取的淋巴提取物中存在的一种序列特异性因子的结合活性。我们扩展了这些研究，以表明该结合活性存在于正常激活的脾B细胞和λ产生细胞中，并通过DNA足迹分析证明包含与SV-40核心增强子11个碱基同源的序列完全保护。我们将这些体外结合研究与使用基因组测序技术分析完整小鼠细胞系中的蛋白质-DNA相互作用进行了比较。我们发现小鼠70z/3细胞系在诱导κ表达后，dna的dm5反应性发生了显著的变化。这些改变发生在作为11个碱基核心序列一部分的鸟氨酸残基上，并且与在组成表达κ的细胞中观察到的相同。这为诱导组织特异性因子与完整的计时蛋白结合提供了直接证据。在完整的染色质中，我们还观察到核心序列的GUA9，3‘的反应性发生了显著变化，这是潜在的二级DNA结构的一部分，并保护了四个残基，这些残基是重链增强器同源区域的一部分。
The large intron of the κ immunoglobulin gene contains a cis-acting enhancer element, which is important in the tissue-specific expression of gene. We have confirmed the binding activity of a sequence-specific factor present in lymphoid extracts derived from cell lines expressing, or induced to express, the κ gene. We have extended these studies to show the binding activity is present in normal activated splenic B cells as well as λproducing cells, and have demonstrated by DNAae footprint analysis fullS protection of a sequence containing the 11 bp homology to the SV-40 core enhancer. We have compared these in vitro binding studies with an analysis protein-DNA interactions in intact murine cell lines using genomic sequencing techniques. We demonstrate significant alterations in DM5 reac tivity of DNA in the murine 70Z/3 cell line after it is induced to κ expression. These alterations occur at gu.anine residues which are part of the 11 bp core sequence, and are identical to those observed in cells constitutively expressing κ. This provides direct evidence for the induced binding of the tissue specific factor to intact chroniatin. In intact chromatin we also observed significant alteration in the reactivity of a gua nine, 3′ of the core sequence, which is part of a potential secondary DNA structure, and protection of four residues that are part of a region homologous to the heavy chain enhancer.