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Cited by 305

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CENP-B box is required for de novo centromere chromatin assembly on human alphoid DNA

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Ohzeki, Jun-ichirou, Nakano, Megumi, Okada, Teruaki, Masumoto, Hiroshi·2002·The Journal of Cell Biology

Abstract

Centromere protein (CENP) B boxes, recognition sequences of CENP-B, appear at regular intervals in human centromeric α-satellite DNA (alphoid DNA). In this study, to determine whether information carried by the primary sequence of alphoid DNA is involved in assembly of functional human centromeres, we created four kinds of synthetic repetitive sequences: modified alphoid DNA with point mutations in all CENP-B boxes, resulting in loss of all CENP-B binding activity; unmodified alphoid DNA containing functional CENP-B boxes; and nonalphoid repetitive DNA sequences with or without functional CENP-B boxes. These four synthetic repetitive DNAs were introduced into cultured human cells (HT1080), and de novo centromere assembly was assessed using the mammalian artificial chromosome (MAC) formation assay. We found that both the CENP-B box and the alphoid DNA sequence are required for de novo MAC formation and assembly of functional centromere components such as CENP-A, CENP-C, and CENP-E. Using the chromatin immunoprecipitation assay, we found that direct assembly of CENP-A and CENP-B in cells with synthetic alphoid DNA required functional CENP-B boxes. To the best of our knowledge, this is the first reported evidence of a functional molecular link between a centromere-specific DNA sequence and centromeric chromatin assembly in humans.

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Metadata

Authors
Ohzeki, J.-I., Nakano, M., Okada, T., & Masumoto, H.
Venue
The Journal of Cell Biology
Published
2002-12-02
Volume
159
Issue
5
Pages
765-775
ISSN
  • 0021-9525 (ISSN-L)
  • 1540-8140
OpenAlex
W2114820564
Cited by
305
References
38
FWCI
5.71
Type
journal article
Language
en
Open access
green

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CoChat. (2026). Citation check: CENP-B box is required for de novo centromere chromatin assembly on human alphoid DNA. https://app.cochat.ai/papers/10.1083/jcb.200207112
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Ohzeki, J.-I., Nakano, M., Okada, T., & Masumoto, H. (2002). CENP-B box is required for de novo centromere chromatin assembly on human alphoid DNA. The Journal of Cell Biology. https://doi.org/10.1083/jcb.200207112

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