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

Kehui Wang; Yuting Liu; Zhouliang Yu; Bo Gu; Jie Hu; Li Huang; Xiao Ge; Lingyi Xu; Mengyu Zhang; Jicheng Zhao; Mingli Hu; Rongrong Le; Qiang Wu; Sheng Ye; Shaorong Gao; Xiaodong Zhang; Rui-Ming Xu; Guohong Li

doi:10.1016/j.celrep.2021.109987

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.

Authors

Kehui Wang¹, Yuting Liu², Zhouliang Yu², Bo Gu³, Jie Hu², Li Huang¹, Xiao Ge⁴, Lingyi Xu⁵, Mengyu Zhang⁶, Jicheng Zhao¹, Mingli Hu², Rongrong Le⁷, Qiang Wu⁴, Sheng Ye⁸, Shaorong Gao⁷, Xiaodong Zhang³, Rui-Ming Xu², Guohong Li⁹

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Cycle*
Centromere
Centromere Protein A / chemistry
Centromere Protein A / genetics
Centromere Protein A / metabolism*
Chromatin / genetics
Chromatin / metabolism
Cullin Proteins / genetics
Cullin Proteins / metabolism*
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism*
Female
Histones / genetics
Histones / metabolism
Male
Mice
Mice, Inbred C57BL
Nucleosomes
Phosphorylation
Proteolysis
Serine / chemistry
Serine / genetics
Serine / metabolism*
Ubiquitin / metabolism
Ubiquitin-Protein Ligase Complexes / genetics
Ubiquitin-Protein Ligase Complexes / metabolism*
Ubiquitination*

Substances

Centromere Protein A
Chromatin
Cullin Proteins
DDB1 protein, human
DNA-Binding Proteins
Histones
Nucleosomes
Ubiquitin
Serine
DDB1 and CUL4 associated factor 11, human
Ubiquitin-Protein Ligase Complexes

Grants and funding

55008737/HHMI/Howard Hughes Medical Institute/United States