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EMBO J. 2002 Oct 1;21(19):5269-80.

Co-localization of centromere activity, proteins and topoisomerase II within a subdomain of the major human X alpha-satellite array.

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  • 1Department of Genetics, University of Cambridge, Downing Street, Cambridge CB2 3EH, UK.

Abstract

Dissection of human centromeres is difficult because of the lack of landmarks within highly repeated DNA. We have systematically manipulated a single human X centromere generating a large series of deletion derivatives, which have been examined at four levels: linear DNA structure; the distribution of constitutive centromere proteins; topoisomerase IIalpha cleavage activity; and mitotic stability. We have determined that the human X major alpha-satellite locus, DXZ1, is asymmetrically organized with an active subdomain anchored approximately 150 kb in from the Xp-edge. We demonstrate a major site of topoisomerase II cleavage within this domain that can shift if juxtaposed with a telomere, suggesting that this enzyme recognizes an epigenetic determinant within the DXZ1 chromatin. The observation that the only part of the DXZ1 locus shared by all deletion derivatives is a highly restricted region of <50 kb, which coincides with the topo isomerase II cleavage site, together with the high levels of cleavage detected, identify topoisomerase II as a major player in centromere biology.

PMID:
12356743
[PubMed - indexed for MEDLINE]
PMCID:
PMC129033
Free PMC Article

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