Neocentromere formation using chromosome engineering

Press release

Chromosome engineering allows the efficient isolation of vertebrate neocentromeres

Wei-Hao Shang, Tetsuya Hori, Nuno M.C. Martins, Atsushi Toyoda, Sadahiko Misu, Norikazu Monma, Ichiro Hiratani, Kazuhiro Maeshima, Kazuho Ikeo, Asao Fujiyama, Hiroshi Kimura, William C. Earnshaw, and Tatsuo Fukagawa
Developmental Cell, 24(6), 635-648, 14 March 2013. DOI: doi:10.1016/j.devcel.2013.02.009

Centromeres are specified by sequence-independent epigenetic mechanisms in most organisms. Rarely, centromere repositioning results in neocentromere formation at ectopic sites. However, the mechanisms governing how and where neocentromeres form are unknown.

Here, we established a chromosome-engineering system in chicken DT40 cells that allowed us to efficiently isolate neocentromere-containing chromosomes.

Neocentromeres appear to be structurally and functionally equivalent to native centromeres. Chromatin immunoprecipitation sequencing (ChIP-seq) analysis with 18 neocentromeres revealed that the centromere-specific histone H3 variant CENP-A occupies an ∼40 kb region at each neocentromere, which has no preference for specific DNA sequence motifs. Furthermore, we found that neocentromeres were not associated with histone modifications H3K9me3, H3K4me2, and H3K36me3 or with early replication timing. Importantly, low but significant levels of CENP-A are detected around endogenous centromeres, which are capable of seeding neocentromere assembly if the centromere core is removed.

Our experimental system provides valuable insights for understanding how neocentromeres form. This was performed by Wei-Hao Shang and Tetsuya Hori (Fukagawa Lab) in collaboration with TRIC of ROIS, Fujimaya Lab. (NIG, NII), Maeshima Lab (NIG), Ikeo Lab. (NIG), Kimura Lab (Osaka U.), and Eranshaw Lab (U. Edinburgh).


Cell division and centromere
During mitosis spindle microtubules capture a special structure of chromosome for faithful chromosome segregation. This structure is called “Kinetochore”. Centromere is a genome region in which kinetochore is formed. Dysfunction of kinetochore results in some diseases including cancer.

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