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Neddylation and deneddylation regulate Cul1 and Cul3 protein accumulation

Nature Cell Biology volume 7, pages 1014–1020 (2005)Cite this article

Abstract

Cullin family proteins organize ubiquitin ligase (E3) complexes to target numerous cellular proteins for proteasomal degradation. Neddylation, the process that conjugates the ubiquitin-like polypeptide Nedd8 to the conserved lysines of cullins, is essential for in vivo cullin-organized E3 activities1,2. Deneddylation, which removes the Nedd8 moiety, requires the isopeptidase activity of the COP9 signalosome (CSN)3,4. Here we show that in cells deficient for CSN activity, cullin1 (Cul1) and cullin3 (Cul3) proteins are unstable, and that to preserve their normal cellular levels, CSN isopeptidase activity is required. We further show that neddylated Cul1 and Cul3 are unstable — as suggested by the evidence that Nedd8 promotes the instability of both cullins — and that the unneddylatable forms of cullins are stable. The protein stability of Nedd8 is also subject to CSN regulation and this regulation depends on its cullin-conjugating ability, suggesting that Nedd8-conjugated cullins are degraded en bloc. We propose that while Nedd8 promotes cullin activation through neddylation, neddylation also renders cullins unstable. Thus, CSN deneddylation recycles the unstable, neddylated cullins into stable, unneddylated ones, and promotes cullin-organized E3 activity in vivo.

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Figure 1: Reduction of Cul1 and Cul3 protein levels in cells harbouring mutations in the CSN complex.
Figure 2: Degradation of Cul1 and Cul3 proteins.
Figure 3: Requirement of CSN deneddylating activity to maintain Cul1 and Cul3 protein stability.
Figure 4: CSN regulation of Cul1 and Cul3 protein levels through a neddylation-dependent mechanism.
Figure 5: Degradation of Nedd8 in CSN5null cells.

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Acknowledgements

The authors thank S. K. Beckendorf, D. A. Chamovitz, G. Cope, R. Desheias and D. Segal for reagents and fly stocks; S. K. Huang and Y. R. Chan for generating the Nedd8 transgenic stocks; K. Deen for editing work; and E. Chang, S. I. Jang, C. H. Lee, T. P. Yao and lab members for helpful comments and discussions. This work was supported by National Science Council, National Health Research Institute, and Academia Sinica of Taiwan to C.T.C. and NHRI MD PhD/DDS PhD Predoctoral Fellowship DD9307N to J.T.W.

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Authors and Affiliations

  1. Institute of Molecular Biology, Academia Sinica, 115, Taipei, Taiwan

    June-Tai Wu, Hsiu-Chen Lin, Yen-Chen Hu & Cheng-Ting Chien

  2. Institute of Molecular Medicine, Medical College, National Taiwan University, 100, Taipei, Taiwan

    June-Tai Wu & Cheng-Ting Chien

  3. Graduate Institute of Life Sciences, National Defense Medical Center, 114, Taipei, Taiwan

    Hsiu-Chen Lin & Cheng-Ting Chien

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Correspondence to Cheng-Ting Chien.

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Wu, JT., Lin, HC., Hu, YC. et al. Neddylation and deneddylation regulate Cul1 and Cul3 protein accumulation. Nat Cell Biol 7, 1014–1020 (2005). https://doi.org/10.1038/ncb1301

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