Chromosome Engineering Allows the Efficient Isolation of Vertebrate Neocentromeres

摘要: Centromeres are specified by sequence-independent epigenetic mechanisms in most organisms. Rarely, centromere repositioning results neocentromere formation at ectopic sites. However, the governing how and where neocentromeres form unknown. Here, we established a chromosome-engineering system chicken DT40 cells that allowed us to efficiently isolate neocentromere-containing chromosomes. Neocentromeres appear be structurally functionally equivalent native centromeres. Chromatin immunoprecipitation sequencing (ChIP-seq) analysis with 18 revealed centromere-specific histone H3 variant CENP-A occupies an ∼40 kb region each neocentromere, which has no preference for specific DNA sequence motifs. Furthermore, found were not associated modifications H3K9me3, H3K4me2, H3K36me3 or early replication timing. Importantly, low but significant levels of detected around endogenous centromeres, capable seeding assembly if core is removed. In summary, our experimental provides valuable insights understanding form.