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Targeting of protein ubiquitination by BTB–Cullin 3–Roc1 ubiquitin ligases

Nature Cell Biology volume 5, pages 1001–1007 (2003)Cite this article

Abstract

The concentrations and functions of many cellular proteins are regulated by the ubiquitin pathway. Cullin family proteins bind with the RING-finger protein Roc1 to recruit the ubiquitin-conjugating enzyme (E2) to the ubiquitin ligase complex (E3). Cul1 and Cul7, but not other cullins, bind to an adaptor protein, Skp1. Cul1 associates with one of many F-box proteins through Skp1 to assemble various SCF–Roc1 E3 ligases that each selectively ubiquitinate one or more specific substrates. Here, we show that Cul3, but not other cullins, binds directly to multiple BTB domains through a conserved amino-terminal domain. In vitro, Cul3 promoted ubiquitination of Caenorhabditis elegans MEI-1, a katanin-like protein whose degradation requires the function of both Cul3 and BTB protein MEL-26. We suggest that in vivo there exists a potentially large number of BCR3 (BTB–Cul3–Roc1) E3 ubiquitin ligases.

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Figure 1: Purification of CUL3 complexes.
Figure 2: BTB domain of KIAA1309 binds to an N-terminal sequence of Cul3.
Figure 3: Cul3 binds to multiple BTB domains.
Figure 4: Cul3 binds to MEL-26 and MEI-1.
Figure 5: MEL26-Cul3–Roc1 promotes MEI-1 ubiquitination in vitro.

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Acknowledgements

We thank the Kazusa DNA Research Institute (Chiba, Japan) for providing KIAA EST clones, S. Ahmed for providing the C. elegans library, N. Zheng for providing Cul3 and Roc1 baculoviruses, and C. McCall, J. McCarville and other members of the Xiong lab for discussion throughout this work and critically reading the manuscript. Y. X. is supported in part by a US Department of Defense Career Development Award. This study is supported by a National Institutes of Health grant to Y.X.

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  1. Department of Biochemistry and Biophysics, Lineberger Comprehensive Cancer Center, and Program in Molecular Biology and Biotechnology, University of North Carolina, Chapel Hill, 27599-7295, NC, USA

    Manabu Furukawa, Yizhou Joseph He, Christoph Borchers & Yue Xiong

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Correspondence to Yue Xiong.

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Furukawa, M., He, Y., Borchers, C. et al. Targeting of protein ubiquitination by BTB–Cullin 3–Roc1 ubiquitin ligases. Nat Cell Biol 5, 1001–1007 (2003). https://doi.org/10.1038/ncb1056

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