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A novel chromatin immunoprecipitation and array (CIA) analysis identifies a 460-kb CENP-A-binding neocentromere DNA
journal contribution
posted on 2001-03-01, 00:00 authored by A W Lo, D J Magliano, M C Sibson, P Kalitsis, Jeffrey CraigJeffrey Craig, K H ChooCentromere protein A (CENP-A) is an essential histone H3-related protein that constitutes the specialized chromatin of an active centromere. It has been suggested that this protein plays a key role in the epigenetic marking and transformation of noncentromeric genomic DNA into functional neocentromeres. Neocentromeres have been identified on more than two-thirds of the human chromosomes, presumably involving different noncentromeric DNA sequences, but it is unclear whether some generalized sequence properties account for these neocentromeric sites. Using a novel method combining chromatin immunoprecipitation and genomic array hybridization, we have identified a 460-kb CENP-A-binding DNA domain of a neocentromere derived from the 20p12 region of an invdup (20p) human marker chromosome. Detailed sequence analysis indicates that this domain contains no centromeric alpha-satellite, classical satellites, or other known pericentric repetitive sequence motifs. Putative gene loci are detected, suggesting that their presence does not preclude neocentromere formation. The sequence is not significantly different from surrounding non-CENP-A-binding DNA in terms of the prevalence of various interspersed repeats and binding sites for DNA-interacting proteins (Topoisomerase II and High-Mobility-Group protein I). Notable variations include a higher AT content similar to that seen in human alpha-satellite DNA and a reduced prevalence of long terminal repeats (LTRs), short interspersed repeats (SINEs), and Alus. The significance of these features in neocentromerization is discussed.
History
Journal
Genome researchVolume
11Issue
3Pagination
448 - 457Publisher
Cold Spring Harbor Laboratory PressLocation
Cold Spring Harbor, N.Y.Publisher DOI
Link to full text
ISSN
1088-9051Language
engPublication classification
C1.1 Refereed article in a scholarly journalCopyright notice
2001, Cold Spring Harbor Laboratory PressUsage metrics
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Alu ElementsAutoantigensBase CompositionCell Line, TransformedCentromereCentromere Protein AChromatinChromosomal Proteins, Non-HistoneChromosome MappingChromosomes, Human, Pair 20DNA-Binding ProteinsGene Expression ProfilingHumansIn Situ Hybridization, FluorescenceMicroscopy, FluorescenceOligonucleotide Array Sequence AnalysisPrecipitin TestsProtein Structure, TertiaryShort Interspersed Nucleotide ElementsScience & TechnologyLife Sciences & BiomedicineBiochemistry & Molecular BiologyBiotechnology & Applied MicrobiologyGenetics & HeredityALPHA-SATELLITE DNATOPOISOMERASE-IIHUMAN CENTROMERE10Q25 NEOCENTROMERESEQUENCE-ANALYSISPROTEIN BINDSHISTONE H3KINETOCHORENUCLEOSOMEORGANIZATION
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