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首页> 外文期刊>Development >Changes in the nuclear deposition of histone H2A variants during pre-implantation development in mice.
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Changes in the nuclear deposition of histone H2A variants during pre-implantation development in mice.

机译：组蛋白H2A变体在小鼠植入前发育过程中核沉积的变化。

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Histone H2A has several variants, and changes in chromatin composition associated with their replacement might involve chromatin structure remodeling. We examined the dynamics of the canonical histone H2A and its three variants, H2A.X, H2A.Z and macroH2A, in the mouse during oogenesis and pre-implantation development when genome remodeling occurs. Immunocytochemistry with specific antibodies revealed that, although H2A and all variants were deposited in the nuclei of full-grown oocytes, only histone H2A.X was abundant in the pronuclei of one-cell embryos after fertilization, in contrast with the low abundance of histone H2A and the absence of H2A.Z. The decline in H2A and the depletion of H2A.Z and macroH2A after fertilization were confirmed using Flag epitope-tagged H2A, H2A.Z and macroH2A transgenic mouse lines. Microinjection experiments with mRNA encoding the Flag-tagged proteins revealed a similar pattern of nuclear incorporation of the H2A variants. Fusion protein experiments using H2A, H2A.Z and macroH2A fused with the C-terminal 23 amino acids of H2A.X showed that the C-terminal amino acids of H2A.X function specifically to target this variant histone into chromatin in embryos after fertilization and that the absence of H2A.Z and macroH2A from the chromatin is required for normal development. These results suggest that global changes in the composition of histone H2A variants in chromatin play a role in genome remodeling after fertilization.
机译：组蛋白H2A具有多种变体，与其替换相关的染色质组成变化可能涉及染色质结构重塑。我们在基因组重塑发生的卵子发生和植入前发育过程中，检查了小鼠中规范组蛋白H2A及其三个变体H2A.X，H2A.Z和macroH2A的动力学。用特异性抗体进行的免疫细胞化学分析显示，尽管H2A和所有变体都沉积在成熟卵母细胞的核中，但受精后单细胞胚胎的前核中只有组蛋白H2A.X丰富，而组蛋白H2A的丰度却低并且没有H2A.Z。使用Flag表位标记的H2A，H2A.Z和macroH2A转基因小鼠品系证实了受精后H2A的下降以及H2A.Z和macroH2A的消耗。用mRNA编码带有Flag标签的蛋白的显微注射实验揭示了H2A变体的核掺入相似模式。使用H2A，H2A.Z和macroH2A与H2A.X的C末端23个氨基酸融合的融合蛋白实验显示，H2A.X的C末端氨基酸可特异性地将这种变体组蛋白靶向受精和胚胎后的染色质。染色质中不存在H2A.Z和macroH2A是正常发育所必需的。这些结果表明，染色质中组蛋白H2A变体组成的整体变化在受精后的基因组重塑中起作用。

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