High-throughput screening of genome fragments bound to differentially acetylated histones
High-throughput screening of genome fragments bound to differentially acetylated histones
复制标题
DOI：
10.1111/j.1365-2443.2004.00804.x
复制
发表时间：
2004-12-01
期刊：
影响因子：
2.1
通讯作者：
Mano, H
中科院分区：
文献类型：
--
作者：
Kaneda, R;Toyota, M;Mano, H
虽然组蛋白的乙酰化-去乙酰化有助于基因表达的调节，但很少有方法可以确定特定细胞或组织中的全基因组组蛋白乙酰化谱。我们现在已经开发了一种全基因组筛选方法，差分染色质扫描（DCS），以分离嵌入组蛋白的基因组片段，这些组蛋白受到差异乙酰化。这种DCS筛选应用于有或没有组蛋白去乙酰化酶（HDAC）活性抑制剂孵育的人胃癌细胞系，结果快速鉴定了250多个基因组片段。有趣的是，许多癌症相关基因被发现是癌细胞中HDAC的靶标，包括肿瘤蛋白73和细胞分裂周期34的基因。组蛋白的这种差异乙酰化也被证明与相应基因的转录活性调节有关。在分离的基因组片段中，94%（32/34）被证实与差异乙酰化组蛋白结合，其中78%（7/9）对应的基因表现出与组蛋白乙酰化水平一致的差异转录活性。凭借其高保真度，DCS方法应该可以快速比较全基因组组蛋白乙酰化谱，并为分子癌变提供新的见解。
Although acetylation-deacetylation of histones contributes to regulation of gene expression, few methods have been available to determine the whole-genome histone acetylation profile in specific cells or tissues. We have now developed a genome-wide screening method, differential chromatin scanning (DCS), to isolate genome fragments embedded in histones subject to differential acetylation. This DCS screening was applied to a human gastric cancer cell line incubated with or without an inhibitor of histone deacetylase (HDAC) activity, resulting in the rapid identification of more than 250 genome fragments. Interestingly, a number of cancer-related genes were revealed to be the targets of HDAC in the cancer cells, including those for tumour protein 73 and cell division cycle 34. Such differential acetylation of histone was also shown to be linked to the regulation of transcriptional activity of the corresponding genes. Among the isolated genome fragments, 94% (32/34) of them were confirmed to be bound to differentially acetylated histones, and the genes corresponding to 78% (7/9) of them exhibited differential transcriptional activity consistent with the level of histone acetylation. With its high fidelity, the DCS method should open a possibility to rapidly compare the genome-wide histone acetylation profiles and to provide novel insights into molecular carcinogenesis.