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A 330 kb CENP-A binding domain and altered replication timing at a human neocentromere
journal contribution
posted on 2001-04-17, 00:00 authored by A W Lo, Jeffrey CraigJeffrey Craig, R Saffery, P Kalitsis, D V Irvine, E Earle, D J Magliano, K H ChooCentromere 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 alpha-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.
History
Journal
EMBO journalVolume
20Issue
8Pagination
2087 - 2096Publisher
European Molecular Biology OrganizationLocation
Oxford, Eng.Publisher DOI
ISSN
0261-4189Language
engPublication classification
C1.1 Refereed article in a scholarly journalCopyright notice
2001, European Molecular Biology OrganizationUsage metrics
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AutoantigensBase CompositionCentromereCentromere Protein AChromosomal Proteins, Non-HistoneChromosomes, Human, Pair 10DNA ReplicationDNA, SatelliteHistonesHumansIn Situ Hybridization, FluorescenceNucleosomesOligonucleotide Array Sequence AnalysisProtein BindingSequence Analysis, DNACENP‐AChromatinNeocentromereReplication TimingScience & TechnologyLife Sciences & BiomedicineBiochemistry & Molecular BiologyCell BiologyCENP-AALPHA-SATELLITE DNACENTROMERIC CHROMATINSEQUENCE-ANALYSIS10Q25 NEOCENTROMEREREPETITIVE DNAPROTEINPATTERNSORGANIZATIONCOMPONENTDISTINCT
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