Abstract
In vertebrates, centromeres lack defined sequences and are thought to be propagated by epigenetic mechanisms involving the incorporation of specialized nucleosomes containing the histone H3 variant centromere protein (CENP)-A. However, the precise mechanisms that target CENP-A to centromeres remain poorly understood. Here, we isolated a multi-subunit complex, which includes the established inner kinetochore components CENP-H and CENP-I, and nine other proteins, from both human and chicken cells. Our analysis of these proteins demonstrates that the CENP-H–I complex can be divided into three functional sub-complexes, each of which is required for faithful chromosome segregation. Interestingly, newly expressed CENP-A is not efficiently incorporated into centromeres in knockout mutants of a subclass of CENP-H–I complex proteins, indicating that the CENP-H–I complex may function, in part, as a marker directing CENP-A deposition to centromeres.
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Acknowledgements
The authors are very grateful to M. Takahashi, K. Suzuki and K. Kita for technical assistance. We thank Y. Mikami and M. Kwon for providing materials and H. Kimura for histone H3–GFP plasmid. This work was supported by Grants-in-Aid for Scientific Research on Priority Areas 'Cancer Cell Biology' and 'Nuclear Dynamics' from the Ministry of Education, Science, Sports and Culture of Japan. This work was also supported by funding from the Ludwig Institute for Cancer Research to A.D., a National Institutes of Health grant to A.D. (R01GM074215–01). I.M.C. is a fellow of the Jane Coffin Childs Memorial Fund for Medical Research. A.D. is a Damon Runyon Scholar supported by the Damon Runyon Cancer Research Foundation (DRS 38–04).
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Okada, M., Cheeseman, I., Hori, T. et al. The CENP-H–I complex is required for the efficient incorporation of newly synthesized CENP-A into centromeres. Nat Cell Biol 8, 446–457 (2006). https://doi.org/10.1038/ncb1396
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DOI: https://doi.org/10.1038/ncb1396
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