Abstract
Cytokinesis is required for faithful division of cytoplasmic components and duplicated nuclei into two daughter cells. Midbody, a protein-dense organelle that forms at the intercellular bridge, is indispensable for successful cytokinesis. However, the regulatory mechanism of cytokinesis at the midbody still remains elusive. Here, we unveil a critical role for NudC-like protein 2 (NudCL2), a co-chaperone of heat shock protein 90 (Hsp90), in cytokinesis regulation by stabilizing regulator of chromosome condensation 2 (RCC2) at the midbody in mammalian cells. NudCL2 localizes at the midbody, and its downregulation results in cytokinesis failure, multinucleation, and midbody disorganization. Using iTRAQ-based quantitative proteomic analysis, we find that RCC2 levels are decreased in NudCL2 knockout (KO) cells. Moreover, Hsp90 forms a complex with NudCL2 to stabilize RCC2, which is essential for cytokinesis. RCC2 depletion mirrors phenotypes observed in NudCL2-downregulated cells. Importantly, ectopic expression of RCC2 rescues the cytokinesis defects induced by NudCL2 deletion, but not vice versa. Together, our data reveal the significance of the NudCL2/Hsp90/RCC2 pathway in cytokinesis at the midbody.
| Original language | English |
|---|---|
| Pages (from-to) | 766-782 |
| Number of pages | 17 |
| Journal | Protein and Cell |
| Volume | 15 |
| Issue number | 10 |
| DOIs | |
| State | Published - Oct 1 2024 |
Bibliographical note
Publisher Copyright:© The Author(s) 2024.
Funding
We acknowledge Yuxin Yin (Peking University Institute of Systems Biomedicine, Beijing, China) for kindly providing the pSA-RCC2 plasmid. We are grateful to Li Liu, Junli Xuan, Yingying Huang, Jiajia Wang (Core Facilities of Zhejiang University School of Medicine), and Maoguang Xue (Institute of Genetics and Department of Genetics of Zhejiang University) for their technical support, Qin Han and Jiansheng Guo (Center of Cryo-Electron Microscopy of Zhejiang University) for their help in confocal laser scanning microscope. We thank Yunkun Lu and Zhensheng Hu (Life Sciences Institute, Zhejiang University) for their help in submitting the mass spectrometry proteomic data to the ProteomeXchange Consortium. We acknowledge Teresa G. Valencak from the College of Animal Sciences of Zhejiang University for her help in refining this manuscript. We thank all the members of Zhou’s lab for the helpful comments and suggestions during the work. This study was supported by the National Natural Science Foundation of China (Nos. 32070709, 32270771, and U21A20197), the National Key Research and Development Program of China (Nos. 2019YFA0802202), and the Higher Education Discipline Innovation Project (also known as the111 Project) (Nos. B13026). This study was supported by the National Natural Science Foundation of China (Nos. 32070709, 32270771, and U21A20197), the National Key Research and Development Program of China (Nos. 2019YFA0802202), and the Higher Education Discipline Innovation Project (also known as the111 Project) (Nos. B13026).
| Funders | Funder number |
|---|---|
| Department of Genetics of Zhejiang University | |
| Zhejiang University | |
| Peking University Institute of Systems Biomedicine, Beijing, China | |
| Zhejiang University School of Medicine | |
| National Key Basic Research and Development Program of China | 2019YFA0802202 |
| National Key Basic Research and Development Program of China | |
| National Natural Science Foundation of China (NSFC) | U21A20197, 32270771, 32070709 |
| National Natural Science Foundation of China (NSFC) | |
| Higher Education Discipline Innovation Project | B13026 |
| Higher Education Discipline Innovation Project |
Keywords
- Hsp90
- NudCL2
- RCC2
- cytokinesis
- midbody
ASJC Scopus subject areas
- Biotechnology
- Biochemistry
- Drug Discovery
- Cell Biology