We have combined in vivo and in vitro approaches to investigate the function of CENP-B, a major protein of human centromeric heterochromatin. Expression of epitope-tagged deletion derivatives of CENP-B in HeLa cells revealed that a single domain less than 158 residues from the amino terminus of the protein is sufficient to localize CENP-B to centromeres. Centromere localization was abolished if as few as 28 amino acids were removed from the amino terminus of CENP-B. The centromere localization signal of CENP-B can function in an autonomous fashion, relocating a fused bacterial enzyme to centromeres. The centromere localization domain of CENP-B specifically binds in vitro to a subset of alpha-satellite DNA monomers. These results suggest that the primary mechanism for localization of CENP-B to centromeres involves the recognition of a DNA sequence found at centromeres. Analysis of the distribution of this sequence in alpha-satellite DNA suggests that CENP-B binding may have profound effects on chromatin structure at centromeres.
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1 March 1992
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March 01 1992
Identification of a subdomain of CENP-B that is necessary and sufficient for localization to the human centromere.
A F Pluta,
A F Pluta
Department of Cell Biology and Anatomy, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
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N Saitoh,
N Saitoh
Department of Cell Biology and Anatomy, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
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I Goldberg,
I Goldberg
Department of Cell Biology and Anatomy, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
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W C Earnshaw
W C Earnshaw
Department of Cell Biology and Anatomy, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
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A F Pluta
Department of Cell Biology and Anatomy, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
N Saitoh
Department of Cell Biology and Anatomy, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
I Goldberg
Department of Cell Biology and Anatomy, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
W C Earnshaw
Department of Cell Biology and Anatomy, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Online ISSN: 1540-8140
Print ISSN: 0021-9525
J Cell Biol (1992) 116 (5): 1081–1093.
Citation
A F Pluta, N Saitoh, I Goldberg, W C Earnshaw; Identification of a subdomain of CENP-B that is necessary and sufficient for localization to the human centromere.. J Cell Biol 1 March 1992; 116 (5): 1081–1093. doi: https://doi.org/10.1083/jcb.116.5.1081
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