@article{ac3def66960d46c8b40e5d74d6a0478d,
title = "CENP-A Ubiquitylation Is Indispensable to Cell Viability",
abstract = "CENP-A is a centromere-specific histone H3 variant that epigenetically determines centromere identity, but how CENP-A is deposited at the centromere remains obscure. We previously reported that CENP-A K124 ubiquitylation, mediated by the CUL4A-RBX1-COPS8 complex, is essential for CENP-A deposition at the centromere. However, a recent report stated that CENP-A K124R mutants show no defects in centromere localization and cell viability. In the present study, we found that EYFP tagging induces additional ubiquitylation of EYFP-CENP-A K124R, which allows the mutant protein to bind to HJURP. Using a previously developed conditional CENP-A knockout system and our CENP-A K124R knockin mutant created by the CRISPR-Cas9 system, we show that the Flag-tagged or untagged CENP-A K124R mutant is lethal. This lethality is rescued by monoubiquitin fusion, indicating that CENP-A ubiquitylation is essential for viability.",
keywords = "CENP-A, centromere, centromere identity, conditional knockout system, epigenetics, kinetochore, mitosis, monoubiquitin, posttranslational modifications (PTMs), ubiquitylation",
author = "Yohei Niikura and Risa Kitagawa and Lei Fang and Katsumi Kitagawa",
note = "Funding Information: We thank Chao-Jun Li at the Model Animal Research Center, Nanjing University for mass spectrometry analysis. We thank Yanmin Dalal, Tatsuo Fukagawa, and current researchers at the Model Animal Research Center, Nanjing University, and Greehey Children's Cancer Research Institute for their helpful discussion, experimental guidance, and reagents. We thank Don W. Cleveland, Daniele Fachinetti, Yanmin Dalal, Minh Bui, Gustavo W. Leone, John Thompson, Lawrence S. Kirschner, Amruta Ashtekar, Ben E. Black, Glennis A. Logsdon, Kenji Tago, and Dawn S. Chandler for their generous gifts of reagents. Y.N. was supported by the 16th Six Big Talent Peaks Fund, Natural Science Foundation of Jiangsu Province, and the Jiangsu Province “Double-First-Class” Construction Fund, and this study was supported by NCI grant R21 CA205659. Conceptualization and Methodology, Y.N. and K.K.; Investigation, Y.N. R.K. and L.F.; Writing – Original Draft, Y.N. and K.K.; Writing – Review & Editing, Y.N. R.K. L.F. and K.K.; Funding Acquisition, Y.N. and K.K.; Total Supervision, K.K. The authors declare no competing interests. Funding Information: We thank Chao-Jun Li at the Model Animal Research Center, Nanjing University for mass spectrometry analysis. We thank Yanmin Dalal, Tatsuo Fukagawa, and current researchers at the Model Animal Research Center, Nanjing University, and Greehey Children{\textquoteright}s Cancer Research Institute for their helpful discussion, experimental guidance, and reagents. We thank Don W. Cleveland, Daniele Fachinetti, Yanmin Dalal, Minh Bui, Gustavo W. Leone, John Thompson, Lawrence S. Kirschner, Amruta Ashtekar, Ben E. Black, Glennis A. Logsdon, Kenji Tago, and Dawn S. Chandler for their generous gifts of reagents. Y.N. was supported by the 16 th Six Big Talent Peaks Fund, Natural Science Foundation of Jiangsu Province , and the Jiangsu Province “Double-First-Class” Construction Fund, and this study was supported by NCI grant R21 CA205659 . Publisher Copyright: {\textcopyright} 2019 Elsevier Inc.",
year = "2019",
month = sep,
day = "23",
doi = "10.1016/j.devcel.2019.07.015",
language = "English (US)",
volume = "50",
pages = "683--689.e6",
journal = "Developmental Cell",
issn = "1534-5807",
publisher = "Cell Press",
number = "6",
}