MAPPING POLYCOMB-REPRESSED DOMAINS IN THE BITHORAX COMPLEX USING IN-VIVO FORMALDEHYDE CROSS-LINKED CHROMATIN
MAPPING POLYCOMB-REPRESSED DOMAINS IN THE BITHORAX COMPLEX USING IN-VIVO FORMALDEHYDE CROSS-LINKED CHROMATIN
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DOI：
10.1016/0092-8674(93)90328-n
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发表时间：
1993-12-17
期刊：
影响因子：
64.5
通讯作者：
PARO, R
中科院分区：
文献类型：
--
作者：
ORLANDO, V;PARO, R
多梳族 (Pc-G) 蛋白负责在果蝇发育过程中保持发育调节因子（如同源异型基因）稳定且遗传性地受到抑制。与异染色质形成中涉及的蛋白质类别的一些相似之处表明，Pc-G 在更高阶染色质水平上发挥其功能。在这里，我们绘制了果蝇组织培养细胞同源双胸复合体 (BX-C) 中 Pc 蛋白的分布。我们详细阐述了一种基于体内甲醛交联技术的方法，该方法可以通过用抗 Pc 抗体对交联染色质进行免疫沉淀来显着富集 Pc 相互作用位点。我们发现 Pc 蛋白定量地覆盖了受抑制的 BX-C 基因的大调控区域。相反，我们发现 Abdominal-B 基因在这些细胞和没有任何结合 Pc 蛋白的区域中活跃。
The Polycomb group (Pc-G) proteins are responsible for keeping developmental regulators, like homeotic genes, stably and inheritably repressed during Drosophila development. Several similarities to a protein class involved in heterochromatin formation suggest that the Pc-G exerts its function at the higher order chromatin level. Here we have mapped the distribution of the Pc protein in the homeotic bithorax complex (BX-C) of Drosophila tissue culture cells. We have elaborated a method, based on the in vivo formaldehyde cross-linking technique, that allows a substantial enrichment for Pc-interacting sites by immunoprecipitation of the cross-linked chromatin with anti-Pc antibodies. We find that the Pc protein quantitatively covers large regulatory regions of repressed BX-C genes. Conversely, we find that the Abdominal-B gene is active in these cells and the region devoid of any bound Pc protein.