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Isolation and Fluorescence Imaging for Single-particle Reconstruction of Chlamydomonas Centrioles
Published on: September 21, 2018
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NudC-like protein 2 restrains centriole amplification by stabilizing HERC2
Min Li1, Xiaoyang Xu1, Jun Zhang2
1Department of Cell Biology and the Cancer Center of the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, 310058, China.
Cell Death & Disease
|August 21, 2019
Summary
Nuclear distribution gene C (NudC)-like protein 2 (NudCL2) stabilizes HECT domain and RCC1-like domain-containing protein 2 (HERC2), preventing centriole amplification. Loss of NudCL2 or HERC2 increases USP33, causing extra centrioles, a defect reversed by USP33 knockdown.
Area of Science:
- Cell biology
- Molecular biology
- Cancer research
Background:
- Centriole duplication ensures one centrosome per cell cycle; errors cause amplification, common in cancer.
- Nuclear distribution gene C (NudC)-like protein 2 (NudCL2) is known to localize to centrosomes, but its function is unclear.
Purpose of the Study:
- To investigate the role of NudCL2 in regulating centrosome function and centriole duplication.
- To elucidate the molecular mechanism by which NudCL2 controls centriole duplication fidelity.
Main Methods:
- CRISPR/Cas9 genome editing and small interfering RNA (siRNA) for gene knockout and depletion.
- Quantitative proteomic analysis to identify protein level changes.
- Co-immunoprecipitation to assess protein interactions.
- Hydroxyurea treatment to induce centriole overduplication.
Main Results:
- NudCL2 knockout or depletion causes significant centriole amplification.
- NudCL2 interacts with and stabilizes the E3 ligase HECT domain and RCC1-like domain-containing protein 2 (HERC2).
- Loss of NudCL2 or HERC2 leads to accumulation of ubiquitin-specific peptidase 33 (USP33), promoting centriole amplification.
- USP33 knockdown reverses centriole amplification in NudCL2 or HERC2 deficient cells.
Conclusions:
- NudCL2 is essential for accurate centriole duplication by stabilizing HERC2.
- This NudCL2-HERC2 axis regulates USP33 levels, providing a mechanism to prevent centriole amplification.
- Disruption of this pathway contributes to the centriole amplification observed in cancers.
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