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USP35 dimer prevents its degradation by E3 ligase CHIP through auto-deubiquitinating activity

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Abstract

Recently, a number of reports on the importance of USP35 in cancer have been published. However, very little is known about the exact mechanism by which USP35 activity is regulated. Here, we show the possible regulation of USP35 activity and the structural specificity affecting its function by analyzing various fragments of USP35. Interestingly, the catalytic domain of USP35 alone does not exhibit deubiquitinating activity; in contrast, the C-terminal domain and insertion region in the catalytic domain is required for full USP35 activity. Additionally, through its C-terminal domain, USP35 forms a homodimer that prevents USP35 degradation. CHIP bound to HSP90 interacts with and ubiquitinates USP35. However, when fully functional USP35 undergoes auto-deubiquitination, which attenuates CHIP-mediated ubiquitination. Finally, USP35 dimer is required for deubiquitination of the substrate Aurora B and regulation of faithful mitotic progression. The properties of USP35 identified in this study are a unique homodimer structure, regulation of deubiquitinating activity through this, and utilization of a novel E3 ligase involved in USP35 auto-deubiquitination, which adds another complexity to the regulation of deubiquitinating enzymes.

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Data availability

All relevant data are available from the corresponding authors upon reasonable request.

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Funding

This work was supported by a National Research Foundation of Korea (NRF) grants funded by the Korean government (MSIP) (2019R1A2C2004052, 2022R1A2C1092294) and Korea Institute of Science and Technology (KIST) Institutional Program (2E32312).

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Authors and Affiliations

  1. Biomedical Research Division, Center for Advanced Biomolecular Recognition, Korea Institute of Science and Technology (KIST), Seoul, 02792, Korea

    Jinyoung Park & Yeojin Kim

  2. Division of Bio‑Medical Science and Technology, KIST‑School, University of Science and Technology (UST), Seoul, 02792, Korea

    Jinyoung Park

  3. Research Resources Division, Technological Convergence Center, Korea Institute of Science and Technology (KIST), Seoul, 02792, Korea

    Sang Chul Shin

  4. Pohang Accelerator Laboratory, Pohang University of Science and Technology, Pohang, 37673, Kyungbuk, Korea

    Kyeong Sik Jin

  5. Biomedical Research Division, Medicinal Materials Research Center, Korea Institute of Science and Technology (KIST), Seoul, 02792, Korea

    Min Joon Lim & Eunice EunKyeong Kim

  6. Department of Life Sciences, College of Life Sciences and Biotechnology, Korea University, Seoul, 02841, Korea

    Yeojin Kim

  7. Graduate School of Pharmaceutical Sciences, College of Pharmacy, Ewha Womans University, Seoul, 03760, Korea

    Eun Joo Song

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  1. Jinyoung Park
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Contributions

JP, EEK and EJS contributed to the study’s conception and design. The most of experiments and analysis were performed by JP. SCS, KSJ, and MJL performed structural analysis and contributed to data collection. YK prepared materials and performed the cell analysis. The first draft of the manuscript was written by JP and SCS. EEK and EJS commented and approved the final manuscript, and conceived and supervised the study.

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Correspondence to Eunice EunKyeong Kim or Eun Joo Song.

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Park, J., Shin, S.C., Jin, K.S. et al. USP35 dimer prevents its degradation by E3 ligase CHIP through auto-deubiquitinating activity. Cell. Mol. Life Sci. 80, 112 (2023). https://doi.org/10.1007/s00018-023-04740-9

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