Division of Life Science, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong SAR, China.
Int J Mol Sci. 2024 Oct 21;25(20):11312. doi: 10.3390/ijms252011312.
Dinoflagellate birefringent chromosomes (BfCs) contain some of the largest known genomes, yet they lack typical nucleosomal micrococcal-nuclease protection patterns despite containing variant core histones. One BfC end interacts with extranuclear mitotic microtubules at the nuclear envelope (NE), which remains intact throughout the cell cycle. Ultrastructural studies, polarized light and fluorescence microscopy, and micrococcal nuclease-resistant profiles (MNRPs) revealed that NE-associated chromosome ends persisted post-mitosis. Histone H3K9me3 inhibition caused S-G delay in synchronous cells, without any effects at G. Differential labeling and nuclear envelope swelling upon decompaction indicate an extension of the inner compartment into telosomal anchorages (TAs). Additionally, limited effects of low-concentration sirtinol on bulk BfCs, coupled with distinct mobility patterns in MNase-digested and psoralen-crosslinked nuclei observed on 2D gels, suggest that telomeric nucleosomes (TNs) are the primary histone structures. The absence of a nucleosomal ladder with cDNA probes, the presence of histone H2A and telomere-enriched H3.3 variants, along with the immuno-localization of H3 variants mainly at the NE further reinforce telomeric regions as the main nucleosomal domains. Cumulative biochemical and molecular analyses suggest that telomeric repeats constitute the major octameric MNRPs that provision chromosomal anchorage at the NE.
甲藻双折射染色体(BfC)含有一些已知的最大基因组,尽管含有变体核心组蛋白,但它们缺乏典型的核小体微球菌核酸酶保护模式。一个 BfC 末端与核膜(NE)上的核外有丝分裂微管相互作用,在整个细胞周期中,NE 保持完整。超微结构研究、偏光和荧光显微镜以及微球菌核酸酶抗性谱(MNRP)表明,与 NE 相关的染色体末端在有丝分裂后仍然存在。组蛋白 H3K9me3 抑制导致同步细胞的 S-G 延迟,而对 G 期没有任何影响。差异标记和核膜肿胀在去致密化后表明内隔室延伸到端粒锚定(TA)。此外,低浓度的 sirtinol 对大量 BfC 的影响有限,以及在二维凝胶上观察到的 MNase 消化和补骨脂素交联核中的明显迁移模式表明,端粒核小体(TNs)是主要的组蛋白结构。cDNA 探针不存在核小体梯,存在组蛋白 H2A 和富含端粒的 H3.3 变体,以及 H3 变体主要在 NE 的免疫定位进一步加强了端粒区域作为主要核小体结构域的地位。累积的生化和分子分析表明,端粒重复构成了主要的八聚体 MNRP,为 NE 处的染色体锚定提供了条件。