Linear ubiquitination of p31comet by HOIP couples cytokine response with mitotic regulation

Yifeng Gao; Qing Yin; Yaser Gamallat; Michael G Grant; Aidan H Snell; Xingxing Shi; Lara N Ulstad; Arshita Singh; Tingan Chen; Joseph O Johnson; Dorina Avram; Lixin Wan

doi:10.1186/s13578-025-01416-8

Linear ubiquitination of p31^comet by HOIP couples cytokine response with mitotic regulation

Cell Biosci. 2025 Jun 3;15(1):75. doi: 10.1186/s13578-025-01416-8.

Authors

Yifeng Gao^#¹, Qing Yin^#¹, Yaser Gamallat¹, Michael G Grant¹, Aidan H Snell¹, Xingxing Shi¹, Lara N Ulstad^{1

2}, Arshita Singh^{1

3}, Tingan Chen⁴, Joseph O Johnson⁴, Dorina Avram⁵, Lixin Wan^{6

7}

Affiliations

¹ Department of Molecular Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, 33612, USA.
² Department of Molecular Biosciences, College of Arts and Sciences, University of South Florida, Tampa, FL, 33620, USA.
³ Department of Chemistry, College of Arts and Sciences, University of South Florida, Tampa, FL, 33620, USA.
⁴ Analytic Microscopy Core Facility, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, 33612, USA.
⁵ Department of Immunology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, 33612, USA.
⁶ Department of Molecular Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, 33612, USA. lixin.wan@moffitt.org.
⁷ Department of Cutaneous Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, 33612, USA. lixin.wan@moffitt.org.

^# Contributed equally.

Abstract

Background: Inflammation and genomic instability are among the hallmarks of human cancer. Proinflammatory cytokines induce DNA damage through the accumulation of reactive oxygen and nitrogen species (RONS), which often leads to base alternations. The link between proinflammatory cytokines and chromosomal instability remains largely elusive.

Results: Here, we report that the mitotic checkpoint protein p31^comet (MAD2L1BP) is modified by linear ubiquitination via the E3 ubiquitin ligase HOIP after cytokine stimulation. HOIP-mediated polyubiquitination of p31^comet occurs on its C-terminal lysine residues. Ubiquitinated p31^comet displays reduced binding to PLK1, which phosphorylates and inactivates p31^comet. Thus HOIP positively regulates p31^comet function. Consistent with this notion, HOIP-deficient cells exhibit prolonged mitotic duration similar to p31^comet knockout. Mitotic defects are also more prevalent in cells without HOIP or p31^comet. Moreover, compared with the cells expressing wild-type p31^comet, cells expressing a ubiquitination-deficient p31^comet mutant take more time to complete the M phase.

Conclusions: Our results together uncover a mechanistic link between the proinflammatory cytokines and the mitotic checkpoint pathways. This molecular switch could be explored as a potential therapeutic target in inflammation-driving or p31^comet overexpressed cancer types.

Keywords: Anaphase-promoting complex; CDC20; HOIP; LUBAC; Mitotic checkpoint complex; P31comet; Proinflammatory cytokine; Spindle assembly checkpoint.

Abstract

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