Next-Generation Sequencing Enables Spatiotemporal Resolution of Human Centromere Replication Timing
Next-Generation Sequencing Enables Spatiotemporal Resolution of Human Centromere Replication Timing
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DOI：
10.3390/genes10040269
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发表时间：
2019-04-01
期刊：
影响因子：
3.5
通讯作者：
Koren, Amnon
中科院分区：
文献类型：
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作者：
Massey, Dashiell J.;Kim, Dongsung;Koren, Amnon
着丝粒通过介导对称的染色体分离，在连续的细胞分裂中保持基因组完整性方面发挥关键作用。着丝粒的重复异染色质性质被认为抑制DNA复制，但也导致其在人类参考基因组组装中的代表性不足。因此，着丝粒已被排除在基因组复制定时分析，留下他们的复制时间未解决。然而，最新的人类参考基因组hg38包括着丝粒序列的模型。为了建立获得着丝粒复制时序曲线的实验要求，我们对来自5个人类细胞系的G（1）期和S期细胞进行测序，并将序列读数与hg38进行比对。我们能够推断DNA复制的着丝粒在每一个细胞系，这表明，着丝粒复制发生在中期至晚期S期的时间配置文件。此外，我们发现，复制时间是更多的细胞系之间的着丝粒区域比预期的变化，复制时间的变异分布在全基因组。这些结果表明，这些潜在的，以及未来的序列模型，使高分辨率的复制研究在着丝粒和其他异染色质区域。
Centromeres serve a critical function in preserving genome integrity across sequential cell divisions, by mediating symmetric chromosome segregation. The repetitive, heterochromatic nature of centromeres is thought to be inhibitory to DNA replication, but has also led to their underrepresentation in human reference genome assemblies. Consequently, centromeres have been excluded from genomic replication timing analyses, leaving their time of replication unresolved. However, the most recent human reference genome, hg38, included models of centromere sequences. To establish the experimental requirements for achieving replication timing profiles for centromeres, we sequenced G(1)- and S-phase cells from five human cell lines, and aligned the sequence reads to hg38. We were able to infer DNA replication timing profiles for the centromeres in each of the five cell lines, which showed that centromere replication occurs in mid-to-late S phase. Furthermore, we found that replication timing was more variable between cell lines in the centromere regions than expected, given the distribution of variation in replication timing genome-wide. These results suggest the potential of these, and future, sequence models to enable high-resolution studies of replication in centromeres and other heterochromatic regions.