Propagation of Centromeric Chromatin Requires Exit from Mitosis

Centromeres direct chromosomal inheritance by nucleating assembly of the kinetochore, a large multiprotein complex required for microtubule attachment during mitosis. Centromere identity in humans is epigenetically determined, with no DNA sequence either necessary or sufficient. A prime candidate fo...

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Published in: The Journal of cell biology Vol. 176; no. 6; pp. 795 - 805
Main Authors: Jansen, Lars E T, Black, Ben E, Foltz, Daniel R, Cleveland, Don W
Format: Journal Article
Language: English
Published: United States Rockefeller University Press 03-12-2007
The Rockefeller University Press
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Summary: Centromeres direct chromosomal inheritance by nucleating assembly of the kinetochore, a large multiprotein complex required for microtubule attachment during mitosis. Centromere identity in humans is epigenetically determined, with no DNA sequence either necessary or sufficient. A prime candidate for the epigenetic mark is assembly into centromeric chromatin of centromere protein A (CENP-A), a histone H3 variant found only at functional centromeres. A new covalent fluorescent pulsechase labeling approach using SNAP tagging has now been developed and is used to demonstrate that CENP-A bound to a mature centromere is quantitatively and equally partitioned to sister centromeres generated during S phase, thereby remaining stably associated through multiple cell divisions. Loading of nascent CENP-A on the megabase domains of replicated centromere DNA is shown to require passage through mitosis but not microtubule attachment. Very surprisingly, assembly and stabilization of new CENP-Acontaining nucleosomes is restricted exclusively to the subsequent G1 phase, demonstrating direct coupling between progression through mitosis and assembly/maturation of the next generation of centromeres.
Bibliography: Correspondence to Don W. Cleveland: dcleveland@ucsd.edu
Abbreviations used in this paper: BG, benzylguanine; CENP-A, centromere protein A; PEG, polyethylene glycol; TMR, tetramethylrhodamine.
ISSN: 0021-9525
1540-8140
DOI: 10.1083/jcb.200701066