Large-scale chromatin fibers of living cells display a discontinuous functional organization
Large-scale chromatin fibers of living cells display a discontinuous functional organization
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活细胞的大规模染色质纤维显示出不连续的功能组织
DOI：
10.1007/s004120000123
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发表时间：
2001
期刊：
影响因子：
1.6
通讯作者：
D. Zink
中科院分区：
文献类型：
--
作者：
Nicolas Sadoni;K. Sullivan;P. Weinzierl;E. Stelzer;D. Zink
摘要。我们研究了活细胞的染色质组织与最近开发的组蛋白和DNA标记方法的组合。用组蛋白H2B-GFP（绿色荧光蛋白）融合蛋白标记核小体DNA，并用高分辨率共聚焦显微镜分析活HeLa细胞的染色质组织。在核周区和核周区染色质被组织成长度为2至8 μ m，直径为300至500 nm的大型纤维。在细胞核内部，我们观察到类似的大规模纤维，但除此之外还有聚焦和弥散形式的组织。与标准标记和检测程序的比较揭示了观察到的染色质组织的主要差异。通过对特定时间复制的DNA进行Cy3-dUTP标记，直接比较组蛋白H2B-GFP分布所揭示的染色质组织与染色质的功能组织。在特定时间复制的DNA区域显示出特有的物理和功能特性。对cy3标记灶的分析显示，它们与所有三种形式的染色质组织（纤维状、局灶性和弥漫性）有关。特别是，cy3标记的病灶出现在大尺度纤维的不连续区域。这些结果表明，大规模的染色质纤维具有不连续的功能特征。
Abstract. We investigated the chromatin organization of living cells with a combination of recently developed approaches for histone and DNA labeling. Nucleosomal DNA was labeled with a histone H2B-GFP (green fluorescent protein) fusion protein and the chromatin organization of living HeLa cells was analyzed by high resolution confocal microscopy. Within the perinuclear and perinucleolar regions chromatin was organized into large-scale fibers of 2 to 8 µm in length and 300 to 500 nm in diameter. Within the nuclear interior we observed similar large-scale fibers, but in addition focal as well as diffuse forms of organization. Comparison with standard labeling and detection procedures revealed major differences in the chromatin organization observed. Chromatin organization revealed by the distribution of histone H2B-GFP was directly compared with the functional organization of chromatin by Cy3-dUTP labeling of DNA replicating at a specific time. DNA regions replicating at a specific time display characteristic physical and functional properties. Analysis of Cy3-labeled foci revealed that they are associated with all three forms of chromatin organization (fibrillar, focal and diffuse). In particular, Cy3-labeled foci appeared as discontinuous regions of large-scale fibers. These results demonstrate that large-scale chromatin fibers have discontinuous functional characteristics.
核超微结构：透射电子显微镜和图像分析。
影响因子：
--
作者：
Belmont,AS
通讯作者：
Belmont,AS