Phosphorylation at Ser68 facilitates DCAF11-mediated ubiquitination and degradation of CENP-A during the cell cycle

  • Cell Rep. 2021 Nov 9;37(6):109987. doi: 10.1016/j.celrep.2021.109987.
Kehui Wang  1 ,  Yuting Liu  2 ,  Zhouliang Yu  2 ,  Bo Gu  3 ,  Jie Hu  2 ,  Li Huang  1 ,  Xiao Ge  4 ,  Lingyi Xu  5 ,  Mengyu Zhang  6 ,  Jicheng Zhao  1 ,  Mingli Hu  2 ,  Rongrong Le  7 ,  Qiang Wu  4 ,  Sheng Ye  8 ,  Shaorong Gao  7 ,  Xiaodong Zhang  3 ,  Rui-Ming Xu  2 ,  Guohong Li  9
Affiliations
  • 1. National Laboratory of Bio-macromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Science, Beijing 100101, China.
  • 2. National Laboratory of Bio-macromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Science, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China.
  • 3. College of Life Sciences, Wuhan University, Wuhan 430072, China.
  • 4. Center for Comparative Biomedicine, Key Laboratory of Systems Biomedicine (Ministry of Education), Institute of Systems Biomedicine, Shanghai Jiao Tong University, Shanghai 200240, China.
  • 5. Department of Biophysics, Department of Pathology of Sir Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310058, China; Department of Biophysics, Institute of Neuroscience, NHC and CAMS Key Laboratory of Medical Neurobiology, Zhejiang University School of Medicine, Hangzhou 310058, China.
  • 6. Tianjin Key Laboratory of Function and Application of Biological Macromolecular Structures, School of Life Sciences, Tianjin University, 92 Weijin Road, Nankai District, Tianjin 300072, China.
  • 7. School of Life Sciences and Technology, Tongji University, Shanghai 200092, China.
  • 8. Tianjin Key Laboratory of Function and Application of Biological Macromolecular Structures, School of Life Sciences, Tianjin University, 92 Weijin Road, Nankai District, Tianjin 300072, China; Life Sciences Institute, Zhejiang University, Hangzhou 310058, China.
  • 9. National Laboratory of Bio-macromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Science, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China. Electronic address: [email protected].
PMID: 34758320 DOI: 10.1016/j.celrep.2021.109987
Abstract

CENP-A (centromeric protein A), a histone H3 variant, specifies centromere identity and is essential to centromere maintenance. Little is known about how protein levels of CENP-A are controlled in mammalian cells. Here, we report that the phosphorylation of CENP-A Ser68 primes the ubiquitin-proteasome-mediated proteolysis of CENP-A during mitotic phase in human cultured cells. We identify two major polyubiquitination sites that are responsible for this phosphorylation-dependent degradation. Substituting the two residues, Lys49 and Lys124, with arginines abrogates proper CENP-A degradation and results in CENP-A mislocalization to non-centromeric regions. Furthermore, we find that DCAF11 (DDB1 and CUL4 associated factor 11/WDR23) is the E3 Ligase that specifically mediates the observed polyubiquitination. Deletion of DCAF11 hampers CENP-A degradation and causes its mislocalization. We conclude that the Ser68 phosphorylation plays an important role in regulating cellular CENP-A homeostasis via DCAF11-mediated degradation to prevent ectopic localization of CENP-A during the cell cycle.