Journal
DEVELOPMENTAL CELL
Volume 24, Issue 6, Pages 635-648Publisher
CELL PRESS
DOI: 10.1016/j.devcel.2013.02.009
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Funding
- Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan
- Cabinet Office, Government of Japan
- Wellcome Trust [073915]
- FCT
- Wellcome Trust Centre for Cell Biology [077707, 092076]
- Precursory Research for Embryonic Science and Technology of the Japan Science and Technology Agency
- MEXT
- Transdisciplinary Research Integration Center (TRIC) of Research Organization of Information and Systems
- Grants-in-Aid for Scientific Research [22370063, 23770235, 23370078] Funding Source: KAKEN
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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 similar to 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 centronnere core is removed. In summary, our experimental system provides valuable insights for understanding how neocentromeres form.
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