Departamento de Bioquimica y Biologia Molecular, Facultad de Ciencias Biologicas, Universidad de Concepcion, Concepcion, Chile.
J Cell Biochem. 2013 Nov;114(11):2569-76. doi: 10.1002/jcb.24605.
One of the most frequent chromosomal translocation found in patients with acute myeloid leukemia (AML) is the t(8;21). This translocation involves the RUNX1 and ETO genes. The breakpoints regions for t(8;21) are located at intron 5 and intron 1 of the RUNX1 and ETO gene respectively. To date, no homologous sequences have been found in these regions to explain their recombination. The breakpoint regions of RUNX1 gene are characterized by the presence of DNasaI hypersensitive sites and topoisomerase II cleavage sites, but no information exists about complementary regions of ETO gene. Here, we report analysis of chromatin structure of ETO breakpoint regions. Chromatin immunoprecipitation (ChIP) were performed with antibodies specific to acetylated histone H3, H4, and total histone H1. Nucleosomal distribution at the ETO locus was evaluated by determining total levels of histone H3. Our data show that in myeloid cells, the breakpoint regions at the ETO gene are enriched in hyperacetylated histone H3 compared to a control region of similar size where no translocations have been described. Moreover, acetylated H4 associates with both the whole ETO breakpoint regions as well as the control intron. Interestingly, we observed no H1 association either at the breakpoint regions or the control region of the ETO gene. Our data indicate that a common chromatin structure enriched in acetylated histones is present in breakpoint regions involved in formation of (8;21) leukemic translocation.
在急性髓系白血病(AML)患者中最常见的染色体易位之一是 t(8;21)。这种易位涉及 RUNX1 和 ETO 基因。t(8;21)的断点区域分别位于 RUNX1 和 ETO 基因的内含子 5 和内含子 1。迄今为止,在这些区域中没有发现同源序列来解释它们的重组。RUNX1 基因的断点区域的特征是存在 DNasaI 超敏位点和拓扑异构酶 II 切割位点,但关于 ETO 基因的互补区域没有信息。在这里,我们报告了 ETO 断点区域染色质结构的分析。使用针对乙酰化组蛋白 H3、H4 和总组蛋白 H1 的特异性抗体进行染色质免疫沉淀(ChIP)。通过确定组蛋白 H3 的总水平来评估 ETO 基因座处的核小体分布。我们的数据表明,在髓样细胞中,与未描述易位的大小相似的对照区域相比,ETO 基因的断点区域富含乙酰化组蛋白 H3。此外,乙酰化 H4 与整个 ETO 断点区域以及对照内含子都相关。有趣的是,我们在 ETO 基因的断点区域或对照区域都没有观察到 H1 结合。我们的数据表明,在涉及(8;21)白血病易位形成的断点区域存在富含乙酰化组蛋白的常见染色质结构。