Abstract
Centromere identity is defined by nucleosomes containing CENP-A, a histone H3 variant. The deposition of CENP-A at centromeres is tightly regulated in a cell-cycle-dependent manner. We previously reported that the spatiotemporal control of centromeric CENP-A incorporation is mediated by the phosphorylation of CENP-A Ser68. However, a recent report argued that Ser68 phosphoregulation is dispensable for accurate CENP-A loading. Here, we report that the substitution of Ser68 of endogenous CENP-A with either Gln68 or Glu68 severely impairs CENP-A deposition and cell viability. We also find that mice harboring the corresponding mutations are lethal. Together, these results indicate that the dynamic phosphorylation of Ser68 ensures cell-cycle-dependent CENP-A deposition and cell viability.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
De Rop, V., Padeganeh, A., and Maddox, P.S. (2012). CENP-A: the key player behind centromere identity, propagation, and kinetochore assembly. Chromosoma 121, 527–538.
Dunleavy, E.M., Roche, D., Tagami, H., Lacoste, N., Ray-Gallet, D., Nakamura, Y., Daigo, Y., Nakatani, Y., and Almouzni-Pettinotti, G. (2009). HJURP is a cell-cycle-dependent maintenance and deposition factor of CENP-A at centromeres. Cell 137, 485–497.
Fachinetti, D., Diego Folco, H., Nechemia-Arbely, Y., Valente, L.P., Nguyen, K., Wong, A.J., Zhu, Q., Holland, A.J., Desai, A., Jansen, L.E. T., et al. (2013). A two-step mechanism for epigenetic specification of centromere identity and function. Nat Cell Biol 15, 1056–1066.
Fachinetti, D., Logsdon, G.A., Abdullah, A., Selzer, E.B., Cleveland, D.W., and Black, B.E. (2017). CENP-A modifications on Ser68 and Lys124 are dispensable for establishment, maintenance, and long-term function of human centromeres. Dev Cell 40, 104–113.
Foltz, D.R., Jansen, L.E.T., Bailey, A.O., Yates Iii, J.R., Bassett, E.A., Wood, S., Black, B.E., and Cleveland, D.W. (2009). Centromere-specific assembly of CENP-a nucleosomes is mediated by HJURP. Cell 137, 472–484.
Fukagawa, T., and Earnshaw, W.C. (2014). The centromere: chromatin foundation for the kinetochore machinery. Dev Cell 30, 496–508.
Holland, A.J., Fachinetti, D., Han, J.S., and Cleveland, D.W. (2012). Inducible, reversible system for the rapid and complete degradation of proteins in mammalian cells. Proc Natl Acad Sci USA 109, E3350–E3357.
Hori, T., Cao, J.H., Nishimura, K., Ariyoshi, M., Arimura, Y., Kurumizaka, H., and Fukagawa, T. (2020). Essentiality of CENP-A depends on its binding mode to HJURP. Cell Rep 33, 108388.
Jansen, L.E.T., Black, B.E., Foltz, D.R., and Cleveland, D.W. (2007). Propagation of centromeric chromatin requires exit from mitosis. J Cell Biol 176, 795–805.
Marzluff, W.F., Wagner, E.J., and Duronio, R.J. (2008). Metabolism and regulation of canonical histone mRNAs: life without a poly(A) tail. Nat Rev Genet 9, 843–854.
Schuh, M., Lehner, C.F., and Heidmann, S. (2007). Incorporation of Drosophila CID/CENP-A and CENP-C into centromeres during early embryonic anaphase. Curr Biol 17, 237–243.
Shelby, R.D., Vafa, O., and Sullivan, K.F. (1997). Assembly of CENP-A into centromeric chromatin requires a cooperative array of nucleosomal DNA contact sites. J Cell Biol 136, 501–513.
Shelby, R.D., Monier, K., and Sullivan, K.F. (2000). Chromatin assembly at kinetochores is uncoupled from DNA replication. J Cell Biol 151, 1113–1118.
Srivastava, S., and Foltz, D.R. (2018). Posttranslational modifications of CENP-A: marks of distinction. Chromosoma 127, 279–290.
Wang, K., Yu, Z., Liu, Y., and Li, G. (2017). Ser68 phosphorylation ensures accurate cell-cycle-dependent CENP-A deposition at centromeres. Dev Cell 40, 5–6.
Wang, K., Liu, Y., Yu, Z., Gu, B., Hu, J., Huang, L., Ge, X., Xu, L., Zhang, M., Zhao, J., et al. (2021). Phosphorylation at Ser68 facilitates DCAF11-mediated ubiquitination and degradation of CENP-A during the cell cycle. Cell Rep 37, 109987.
Xiong, C., Wen, Z., Yu, J., Chen, J., Liu, C.P., Zhang, X., Chen, P., Xu, R. M., Li, G. (2018). UBN1/2 of HIRA complex is responsible for recognition and deposition of H3.3 at cis-regulatory elements of genes in mouse ES cells. BMC Biol 16, 110.
Yu, Z., Zhou, X., Wang, W., Deng, W., Fang, J., Hu, H., Wang, Z., Li, S., Cui, L., Shen, J., et al. (2015). Dynamic phosphorylation of CENP-A at Ser68 orchestrates its cell-cycle-dependent deposition at centromeres. Dev Cell 32, 68–81.
Acknowledgements
This work was supported by the Ministry of Science and Technology of China (2017YFA0504202 and 2019YFA0508903), the National Natural Science Foundation of China (31991161 and 32070604), the Beijing Municipal Science and Technology Committee (Z201100005320013), and HHMI International Research Scholar grant (55008737).
Author information
Authors and Affiliations
Corresponding authors
Additional information
Compliance and ethics The author(s) declare that they have no conflict of interest.
Electronic supplementary material
Rights and permissions
About this article
Cite this article
Liu, Y., Wang, K., Huang, L. et al. Ser68 phosphoregulation is essential for CENP-A deposition, centromere function and viability in mice. Sci. China Life Sci. 65, 1881–1889 (2022). https://doi.org/10.1007/s11427-021-2077-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11427-021-2077-1