A 330 kb CENP-A binding domain and altered replication timing at a human neocentromere

Anthony W I Lo, Jeffrey M. Craig, Richard Saffery, Paul Kalitsis, Danielle V. Irvine, Elizabeth Earle, Dianna J. Magliano, K. H Andy Choo

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120 Citations (Scopus)


Centromere protein A (CENP-A) is an essential centromere-specific histone H3 homologue. Using combined chromatin immunoprecipitation and DNA array analysis, we have defined a 330 kb CENP-A binding domain of a 10q25.3 neocentromere found on the human marker chromosome mardel(10). This domain is situated adjacent to the 80 kb region identified previously as the neocentromere site through lower-resolution immunofluorescence/FISH analysis of metaphase chromosomes. The 330 kb CENP-A binding domain shows a depletion of histone H3, providing evidence for the replacement of histone H3 by CENP-A within centromere-specific nucleosomes. The DNA within this domain has a high AT-content comparable to that of α-satellite, a high prevalence of LINEs and tandem repeats, and fewer SINEs and potential genes than the surrounding region. FISH analysis indicates that the normal 10q25.3 genomic region replicates around mid-S phase. Neocentromere formation is accompanied by a replication time lag around but not within the CENP-A binding region, with this lag being significantly more prominent to one side. The availability of fully sequenced genomic markers makes human neocentromeres a powerful model for dissecting the functional domains of complex higher eukaryotic centromeres.

Original languageEnglish
Pages (from-to)2087-2096
Number of pages10
JournalThe EMBO Journal
Issue number8
Publication statusPublished - 17 Apr 2001
Externally publishedYes


  • CENP-A
  • Centromere
  • Chromatin
  • Neocentromere
  • Replication timing

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