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首页> 外文期刊>Molecular and Cellular Biology >Human immunoglobulin kappa gene enhancer: chromatin structure analysis at high resolution.
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Human immunoglobulin kappa gene enhancer: chromatin structure analysis at high resolution.

机译：人免疫球蛋白κ基因增强子：高分辨率的染色质结构分析。

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The murine immunoglobulin kappa gene enhancer has previously been found to coincide with a region of altered chromatin structure reflected in a DNase I hypersensitivity site detectable on Southern blots of B-cell DNA. We examined the chromatin structure of the homologous region of human DNA using the high-resolution electroblotting method originally developed for genomic sequence analysis by G. Church and W. Gilbert (Proc. Natl. Acad. Sci. USA 81:1991-1995, 1984). Analysis of DNA isolated from cells treated in vivo with dimethyl sulfate revealed two B-cell-specific sites of enhanced guanine methylation. Both sites are located within perfect inverted repeats theoretically capable of forming cruciform structures; one of these repeats overlaps an enhancer core sequence. No enhancement or protection of guanine methylation was observed within sequences similar to sites of altered methylation previously described in the immunoglobulin heavy-chain enhancer. Treatment of isolated nuclei with DNase I or a variety of restriction endonucleases defined a B-cell-specific approximately 0.25-kilobase region of enhanced nuclease susceptibility similar to that observed in the murine kappa enhancer. The 130-base-pair DNA segment that shows high sequence conservation between human, mouse, and rabbit DNAs lies at the 5' end of the nuclease-susceptible region.
机译：先前已经发现鼠免疫球蛋白κ基因增强子与在B细胞DNA的Southern印迹上可检测到的DNase I超敏位点所反映的染色质结构改变的区域一致。我们使用最初由G.Church和W.Gilbert（Proc.Natl.Acad.Sci.USA 81：1991-1995，1984年为基因组序列分析开发的高分辨率电印迹方法）检测了人类DNA同源区域的染色质结构。 ）。对从体内用硫酸二甲酯处理过的细胞中分离的DNA的分析显示，鸟嘌呤甲基化增强了两个B细胞特异性位点。两个位点均位于理论上能够形成十字形结构的完美反向重复序列内;这些重复序列之一与增强子核心序列重叠。在类似于先前在免疫球蛋白重链增强子中描述的甲基化改变位点的序列内，未观察到鸟嘌呤甲基化的增强或保护。用DNase I或多种限制性核酸内切酶处理分离出的细胞核，可确定B细胞特异性的约0.25碱基碱基的增强的核酸酶敏感性，与在鼠Kappa增强子中观察到的相似。显示人类，小鼠和兔子DNA之间具有高度序列保守性的130个碱基对的DNA片段位于核酸酶敏感区的5'端。

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