1] Howard Hughes Medical Institute, Boston, Massachusetts, USA. [2] Program in Cellular and Molecular Medicine, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, USA. [3] Division of Hematology/Oncology, Department of Pediatrics, Boston Children's Hospital, Boston, Massachusetts, USA. [4] Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
1] Howard Hughes Medical Institute, Boston, Massachusetts, USA. [2] Program in Cellular and Molecular Medicine, Boston Children's Hospital, Harvard Medical School, Boston, Massachusetts, USA. [3] Division of Hematology/Oncology, Department of Pediatrics, Boston Children's Hospital, Boston, Massachusetts, USA. [4] Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA. [5] Harvard Stem Cell Institute, Boston, Massachusetts, USA.
Nat Struct Mol Biol. 2014 Jul;21(7):609-16. doi: 10.1038/nsmb.2839. Epub 2014 Jun 8.
Packaging of DNA into nucleosomes not only helps to store genetic information but also creates diverse means for regulating DNA-templated processes. Attempts to reveal additional functions of the nucleosome have been unsuccessful, owing to cell lethality caused by nucleosome deletion. Taking advantage of the mammalian fertilization process, in which sperm DNA assembles into nucleosomes de novo, we generated nucleosome-depleted (ND) paternal pronuclei by depleting maternal histone H3.3 or its chaperone HIRA in mouse zygotes. We found that the ND pronucleus forms a nuclear envelope devoid of nuclear pore complexes (NPCs). Loss of NPCs is accompanied by defective localization of ELYS, a nucleoporin essential for NPC assembly, to the nuclear rim. Interestingly, tethering ELYS to the nuclear rim of the ND nucleus rescues NPC assembly. Our study thus demonstrates that nucleosome assembly is a prerequisite for NPC assembly during paternal pronuclear formation.
将 DNA 包装成核小体不仅有助于储存遗传信息,还为调控 DNA 模板过程创造了多样化的手段。由于核小体缺失会导致细胞死亡,因此尝试揭示核小体的其他功能一直未能成功。利用哺乳动物受精过程中精子 DNA 从头组装成核小体的特点,我们通过耗尽母源性组蛋白 H3.3 或其伴侣蛋白 HIRA,在小鼠受精卵中生成了核小体缺失(ND)的雄性原核。我们发现 ND 原核形成了一种没有核孔复合物(NPCs)的核膜。NPCs 的缺失伴随着核孔蛋白 ELYS 的定位缺陷,ELYS 是 NPC 组装所必需的核孔蛋白。有趣的是,将 ELYS 锚定在 ND 核的核膜边缘可以挽救 NPC 的组装。因此,我们的研究表明,核小体组装是雄性原核形成过程中 NPC 组装的必要条件。