Journal
JOURNAL OF CELL BIOLOGY
Volume 176, Issue 6, Pages 795-805Publisher
ROCKEFELLER UNIV PRESS
DOI: 10.1083/jcb.200701066
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Funding
- NIGMS NIH HHS [R37 GM029513, R01 GM029513, GM74150, R01 GM074150, GM29513] Funding Source: Medline
- NINDS NIH HHS [P30 NS047101, NS047101] Funding Source: Medline
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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 pulse-chase 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-A containing 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.
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