Packiaraj Jenika, Thakur Jitendra
Department of Biology, Emory University, 1510 Clifton Rd, Atlanta, GA 30322.
bioRxiv. 2023 Jul 19:2023.07.18.549612. doi: 10.1101/2023.07.18.549612.
Centromeres are essential for faithful chromosome segregation during mitosis and meiosis. However, the organization of satellite DNA and chromatin at mouse centromeres and pericentromeres is poorly understood due to the challenges of sequencing and assembling repetitive genomic regions. Using recently available PacBio long-read sequencing data from the C57BL/6 strain and chromatin profiling, we found that contrary to the previous reports of their highly homogeneous nature, centromeric and pericentromeric satellites display varied sequences and organization. We find that both centromeric minor satellites and pericentromeric major satellites exhibited sequence variations within and between arrays. While most arrays are continuous, a significant fraction is interspersed with non-satellite sequences, including transposable elements. Additionally, we investigated CENP-A and H3K9me3 chromatin organization at centromeres and pericentromeres using Chromatin immunoprecipitation sequencing (ChIP-seq). We found that the occupancy of CENP-A and H3K9me3 chromatin at centromeric and pericentric regions, respectively, is associated with increased sequence abundance and homogeneity at these regions. Furthermore, the transposable elements at centromeric regions are not part of functional centromeres as they lack CENP-A enrichment. Finally, we found that while H3K9me3 nucleosomes display a well-phased organization on major satellite arrays, CENP-A nucleosomes on minor satellite arrays lack phased organization. Interestingly, the homogeneous class of major satellites phase CENP-A and H3K27me3 nucleosomes as well, indicating that the nucleosome phasing is an inherent property of homogeneous major satellites. Overall, our findings reveal that house mouse centromeres and pericentromeres, which were previously thought to be highly homogenous, display significant diversity in satellite sequence, organization, and chromatin structure.
着丝粒对于有丝分裂和减数分裂过程中染色体的准确分离至关重要。然而,由于对重复基因组区域进行测序和组装存在挑战,小鼠着丝粒和着丝粒周围区域的卫星DNA和染色质的组织情况仍知之甚少。利用最近获得的C57BL/6品系的PacBio长读长测序数据和染色质分析,我们发现,与之前关于其高度同质性质的报道相反,着丝粒和着丝粒周围的卫星显示出不同的序列和组织。我们发现,着丝粒小卫星和着丝粒周围大卫星在阵列内部和之间都表现出序列变异。虽然大多数阵列是连续的,但很大一部分穿插着非卫星序列,包括转座元件。此外,我们使用染色质免疫沉淀测序(ChIP-seq)研究了着丝粒和着丝粒周围区域的CENP-A和H3K9me3染色质组织。我们发现,CENP-A和H3K9me3染色质分别在着丝粒和着丝粒周围区域的占据与这些区域序列丰度和同质性的增加有关。此外,着丝粒区域的转座元件不是功能性着丝粒的一部分,因为它们缺乏CENP-A富集。最后,我们发现,虽然H3K9me3核小体在大卫星阵列上呈现出良好的相位组织,但小卫星阵列上的CENP-A核小体缺乏相位组织。有趣的是,同质类别的大卫星也使CENP-A和H3K27me3核小体相位化,这表明核小体相位化是同质大卫星的固有特性。总体而言,我们的研究结果表明,以前被认为高度同质的家鼠着丝粒和着丝粒周围区域在卫星序列、组织和染色质结构上表现出显著的多样性。