Assembly in G1 phase and long-term stability are unique intrinsic features of CENP-A nucleosomes
- 1 April 2013
- journal article
- Published by American Society for Cell Biology (ASCB) in Molecular Biology of the Cell
- Vol. 24 (7), 923-932
- https://doi.org/10.1091/mbc.e13-01-0034
Abstract
Centromeres are the site of kinetochore formation during mitosis. Centromere protein A (CENP-A), the centromere-specific histone H3 variant, is essential for the epigenetic maintenance of centromere position. Previously we showed that newly synthesized CENP-A is targeted to centromeres exclusively during early G1 phase and is subsequently maintained across mitotic divisions. Using SNAP-based fluorescent pulse labeling, we now demonstrate that cell cycle–restricted chromatin assembly at centromeres is unique to CENP-A nucleosomes and does not involve assembly of other H3 variants. Strikingly, stable retention is restricted to the CENP-A/H4 core of the nucleosome, which we find to outlast general chromatin across several cell divisions. We further show that cell cycle timing of CENP-A assembly is independent of centromeric DNA sequences and instead is mediated by the CENP-A targeting domain. Unexpectedly, this domain also induces stable transmission of centromeric nucleosomes, independent of the CENP-A deposition factor HJURP. This demonstrates that intrinsic properties of the CENP-A protein direct its cell cycle–restricted assembly and induces quantitative mitotic transmission of the CENP-A/H4 nucleosome core, ensuring long-term stability and epigenetic maintenance of centromere position.Keywords
This publication has 50 references indexed in Scilit:
- Analysis of Protein Turnover by Quantitative SNAP‐Based Pulse‐Chase ImagingCurrent Protocols in Cell Biology, 2012
- Labeling of fusion proteins with synthetic fluorophores in live cellsProceedings of the National Academy of Sciences of the United States of America, 2004
- Human centromere repositioning “in progress”Proceedings of the National Academy of Sciences of the United States of America, 2004
- Histone H3 variants specify modes of chromatin assemblyProceedings of the National Academy of Sciences of the United States of America, 2002
- A general method for the covalent labeling of fusion proteins with small molecules in vivoNature Biotechnology, 2002
- Histone mRNA expression: multiple levels of cell cycle regulation and important developmental consequencesCurrent Opinion in Cell Biology, 2002
- CENP-B box is required for de novo centromere chromatin assembly on human alphoid DNAThe Journal of cell biology, 2002
- HIRA Is Critical for a Nucleosome Assembly Pathway Independent of DNA SynthesisMolecular Cell, 2002
- Conserved Organization of Centromeric Chromatin in Flies and HumansDevelopmental Cell, 2002
- Human centromere protein A (CENP-A) can replace histone H3 in nucleosome reconstitution in vitroProceedings of the National Academy of Sciences of the United States of America, 2000