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首页> 外文期刊>Molecular and Cellular Biology >Long-Range Interactions between Three Transcriptional Enhancers, Active Vκ Gene Promoters, and a 3′ Boundary Sequence Spanning 46 Kilobases
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Long-Range Interactions between Three Transcriptional Enhancers, Active Vκ Gene Promoters, and a 3′ Boundary Sequence Spanning 46 Kilobases

机译：三个转录增强子，活性Vκ基因启动子和跨越46个碱基的3'边界序列之间的远程相互作用。

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The mouse immunoglobulin kappa (Igκ) gene contains an intronic enhancer and two enhancers downstream of its transcription unit. Using chromosome conformation capture technology, we demonstrate that rearranged and actively transcribed Igκ alleles in MPC-11 plasmacytoma cells exhibit mutual interactions over 22 kb between these three enhancers and Vκ gene promoters. In addition, the 5′ region of the active transcription unit exhibits a continuum of interactions with downstream chromatin segments. We also observe interactions between Ei and E3′ with 3′ boundary sequences 24 kb downstream of Ed, adjacent to a neighboring housekeeping gene. Very similar interactions between the enhancers are also exhibited by normal B cells isolated from mouse splenic tissue but not by germ line transcriptionally inactive alleles of T cells or P815 mastocytoma cells, which exhibit a seemingly linear chromatin organization. These results fit a looping mechanism for enhancer function like in the β-globin locus and suggest a dynamic modulation of the spatial organization of the active Igκ locus. Chromatin immunoprecipitation experiments reveal that the interacting Igκ gene cis-acting sequences are associated with AP-4, E47, and p65NF-κB, potential protein candidates that may be responsible for initiating and/or maintaining the formation of these higher-order complexes. However, S107 plasmacytoma cells that lack NF-κB still exhibit mutual interactions between the Igκ gene enhancers.
机译：小鼠免疫球蛋白κ（Igκ）基因在其转录单位下游包含一个内含子增强子和两个增强子。使用染色体构象捕获技术，我们证明了MPC-11浆细胞瘤细胞中重排和主动转录的Igκ等位基因在这三个增强子和Vκ基因启动子之间的相互作用超过22 kb。另外，活性转录单位的5'区域表现出与下游染色质片段的连续相互作用。我们还观察到Ei和E3'与位于Ed下游24 kb的3'边界序列之间的相互作用，与相邻的管家基因相邻。从小鼠脾脏组织分离的正常B细胞​​也表现出增强子之间非常相似的相互作用，但T细胞或P815肥大细胞瘤细胞的生殖系转录失活等位基因却没有，它们表现出线性的染色质组织。这些结果适合增强剂功能的循环机制，例如在β珠蛋白基因座中，并提示动态调节活性Igκ基因座的空间组织。染色质免疫沉淀实验表明，相互作用的Igκ基因 cis 作用序列与AP-4，E47和p65NF-κB有关，AP-4，E47和p65NF-κB可能是潜在的蛋白候选物，可能负责启动和/或维持其形成。这些高阶复合体。然而，缺乏NF-κB的S107浆细胞瘤细胞仍表现出Igκ基因增强子之间的相互作用。

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