The BTB protein MEL-26 is a substrate-specific adaptor of the CUL-3 ubiquitin-ligase (original) (raw)

Nature volume 425, pages 311–316 (2003)Cite this article

Abstract

Many biological processes, such as development and cell cycle progression are tightly controlled by selective ubiquitin-dependent degradation of key substrates. In this pathway, the E3-ligase recognizes the substrate and targets it for degradation by the 26S proteasome. The SCF (Skp1–Cul1–F-box) and ECS (Elongin C–Cul2–SOCS box) complexes are two well-defined cullin-based E3-ligases1,2,3. The cullin subunits serve a scaffolding function and interact through their C terminus with the RING-finger-containing protein Hrt1/Roc1/Rbx1, and through their N terminus with Skp1 or Elongin C, respectively. In Caenorhabditis elegans, the ubiquitin-ligase activity of the CUL-3 complex is required for degradation of the microtubule-severing protein MEI-1/katanin at the meiosis-to-mitosis transition4. However, the molecular composition of this cullin-based E3-ligase is not known. Here we identified the BTB-containing protein MEL-26 as a component required for degradation of MEI-1 in vivo. Importantly, MEL-26 specifically interacts with CUL-3 and MEI-1 in vivo and in vitro, and displays properties of a substrate-specific adaptor. Our results suggest that BTB-containing proteins may generally function as substrate-specific adaptors in Cul3-based E3-ubiquitin ligases.

This is a preview of subscription content, access via your institution

Access options

Subscribe to this journal

Receive 51 print issues and online access

$199.00 per year

only $3.90 per issue

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Additional access options:

Similar content being viewed by others

References

  1. Schulman, B. A. et al. Insights into SCF ubiquitin ligases from the structure of the Skp1-Skp2 complex. Nature 408, 381–386 (2000)
    Article ADS CAS PubMed Google Scholar
  2. Zheng, N. et al. Structure of the Cul1-Rbx1-Skp1-F boxSkp2 SCF ubiquitin ligase complex. Nature 416, 703–709 (2002)
    Article ADS CAS PubMed Google Scholar
  3. Stebbins, C. E., Kaelin, W. G. Jr & Pavletich, N. P. Structure of the VHL-ElonginC-ElonginB complex: implications for VHL tumor suppressor function. Science 284, 455–461 (1999)
    Article ADS CAS PubMed Google Scholar
  4. Pintard, L. et al. Neddylation and deneddylation of CUL-3 is required to target MEI-1/Katanin for degradation at the meiosis-to-mitosis transition in C. elegans. Curr. Biol. 13, 911–921 (2003)
    Article CAS PubMed Google Scholar
  5. Zollman, S., Godt, D., Prive, G. G., Couderc, J. L. & Laski, F. A. The BTB domain, found primarily in zinc finger proteins, defines an evolutionarily conserved family that includes several developmentally regulated genes in Drosophila. Proc. Natl Acad. Sci. USA 91, 10717–10721 (1994)
    Article ADS CAS PubMed PubMed Central Google Scholar
  6. Kurz, T. et al. Cytoskeletal regulation by the Nedd8 ubiquitin-like protein modification pathway. Science 295, 1294–1298 (2002)
    Article ADS CAS PubMed Google Scholar
  7. Clark-Maguire, S. & Mains, P. E. Localization of the mei-1 gene product of Caenorhabditis elegans, a meiotic-specific spindle component. J. Cell Biol. 126, 199–209 (1994)
    Article CAS PubMed Google Scholar
  8. Srayko, M., Buster, D. W., Bazirgan, O. A., McNally, F. J. & Mains, P. E. MEI-1/MEI-2 katanin-like microtubule severing activity is required for Caenorhabditis elegans meiosis. Genes Dev. 14, 1072–1084 (2000)
    CAS PubMed PubMed Central Google Scholar
  9. Mains, P. E., Kemphues, K. J., Sprunger, S. A., Sulston, I. A. & Wood, W. B. Mutations affecting the meiotic and mitotic divisions of the early Caenorhabditis elegans embryo. Genetics 126, 593–605 (1990)
    CAS PubMed PubMed Central Google Scholar
  10. Dow, M. R. & Mains, P. E. Genetic and molecular characterization of the Caenorhabditis elegans gene, mel-26, a postmeiotic negative regulator of mei-1, a meiotic-specific spindle component. Genetics 150, 119–128 (1998)
    CAS PubMed PubMed Central Google Scholar
  11. Clark-Maguire, S. & Mains, P. E. mei-1, a gene required for meiotic spindle formation in Caenorhabditis elegans, is a member of a family of ATPases. Genetics 136, 533–546 (1994)
    CAS PubMed PubMed Central Google Scholar
  12. Salama, S. R., Hendricks, K. B. & Thorner, J. G1 cyclin degradation: the PEST motif of yeast Cln2 is necessary, but not sufficient, for rapid protein turnover. Mol. Cell. Biol. 14, 7953–7966 (1994)
    Article CAS PubMed PubMed Central Google Scholar
  13. Galan, J. M. & Peter, M. Ubiquitin-dependent degradation of multiple F-box proteins by an autocatalytic mechanism. Proc. Natl Acad. Sci. USA 96, 9124–9129 (1999)
    Article ADS CAS PubMed PubMed Central Google Scholar
  14. Wirbelauer, C. et al. The F-box protein Skp2 is a ubiquitylation target of a Cul1-based core ubiquitin ligase complex: evidence for a role of Cul1 in the suppression of Skp2 expression in quiescent fibroblasts. EMBO J. 19, 5362–5375 (2000)
    Article CAS PubMed PubMed Central Google Scholar
  15. Zhou, P. & Howley, P. M. Ubiquitination and degradation of the substrate recognition subunits of SCF ubiquitin-protein ligases. Mol. Cell 2, 571–580 (1998)
    Article CAS PubMed Google Scholar
  16. Wolf, D. A., McKeon, F. & Jackson, P. K. F-box/WD-repeat proteins pop1p and Sud1p/Pop2p form complexes that bind and direct the proteolysis of cdc18p. Curr. Biol. 9, 373–376 (1999)
    Article CAS PubMed Google Scholar
  17. Seibert, V. et al. Combinatorial diversity of fission yeast SCF ubiquitin ligases by homo- and heterooligomeric assemblies of the F-box proteins Pop1p and Pop2p. BMC Biochem. 3, 1–15 (2002)
    Article Google Scholar
  18. Robinson, D. N. & Cooley, L. Drosophila kelch is an oligomeric ring canal actin organizer. J. Cell Biol. 138, 799–810 (1997)
    Article CAS PubMed PubMed Central Google Scholar
  19. Soltysik-Espanola, M. et al. Characterization of Mayven, a novel actin-binding protein predominantly expressed in brain. Mol. Biol. Cell 10, 2361–2375 (1999)
    Article CAS PubMed PubMed Central Google Scholar
  20. Bomont, P. et al. The gene encoding gigaxonin, a new member of the cytoskeletal BTB/kelch repeat family, is mutated in giant axonal neuropathy. Nature Genet. 26, 370–374 (2000)
    Article CAS PubMed Google Scholar
  21. Brenner, S. The genetics of Caenorhabditis elegans. Genetics 77, 71–94 (1974)
    CAS PubMed PubMed Central Google Scholar
  22. Ausubel, F. M. et al. Current Protocols in Molecular Biology (Greene Publishing Associates and Wiley-Interscience, New York, 1991)
    Google Scholar
  23. Harper, J. W., Adami, G. R., Wei, N., Keyomarsi, K. & Elledge, S. J. The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases. Cell 75, 805–816 (1993)
    Article CAS PubMed Google Scholar
  24. Breeden, L. K. N. Regulation of the yeast HO gene. Cold Spring Harb. Symp. Quant. Biol. 50, 643–650 (1985)
    Article CAS PubMed Google Scholar
  25. Altschul, S. F. et al. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 25, 3389–3402 (1997)
    Article CAS PubMed PubMed Central Google Scholar
  26. Thompson, J. D., Higgins, D. G. & Gibson, T. J. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22, 4673–4680 (1994)
    Article CAS PubMed PubMed Central Google Scholar
  27. Xu, L. et al. BTB proteins are substrate-specific adaptors in an SCF-like modular ubiquitin ligase containing CUL-3. Nature advance online publication; 3 September 2003 (doi:10.1038/nature01985)

Download references

Acknowledgements

We are grateful to the C. elegans Genetics Centre (funded by the NIH National Centre for Research Resources) for providing strains. We thank R. Fischer for generation of the monoclonal anti-MEL-26 antibody, P. Gönczy for introducing L.P. to C. elegans and for sharing material and reagents, P. Weissert for help with worm liquid cultures, J. M. Bellanger and I. Sumara for suggestions, P. Wiget for help with microscopy, and P. Gönczy for critical reading of the manuscript. L.P. was supported by a Long-Term Fellowship from the Federation of European Biochemical Societies (FEBS) and a Fellowship from Roche, T.K. by a predoctoral fellowship from the American Heart Association, J.H.W. by an NIH Molecular Biology Training Grant, P.E.M. by grants from the Canadian Institutes of Health Research and the Alberta Heritage Foundation for Medical Research, B.B. by the NIH and M.P. by the ETHZ and the Swiss National Science Foundation.

Author information

Author notes

  1. Martin Srayko
    Present address: Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307, Dresden, Germany
  2. Lionel Pintard and John H. Willis: These authors contributed equally to this work

Authors and Affiliations

  1. Institute of Biochemistry ETH, Hönggerberg 8093, Zürich, Switzerland
    Lionel Pintard, Sarah Glaser & Matthias Peter
  2. Institute of Molecular Biology, University of Oregon, Eugene, Oregon, 97403, USA
    John H. Willis, Thimo Kurz & Bruce Bowerman
  3. Department of Medical Genetics and Microbiology, Samuel Lunenfeld Research Institute, Mt Sinai Hosp., Univ. Toronto, 600 University Ave., Toronto, Ontario, M5G1X5, Canada
    Andrew Willems & Mike Tyers
  4. Genes and Development Research Group and Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, Alberta, T2N 4N1, Canada
    Jacque-Lynne F. Johnson, Martin Srayko & Paul E. Mains

Authors

  1. Lionel Pintard
    You can also search for this author inPubMed Google Scholar
  2. John H. Willis
    You can also search for this author inPubMed Google Scholar
  3. Andrew Willems
    You can also search for this author inPubMed Google Scholar
  4. Jacque-Lynne F. Johnson
    You can also search for this author inPubMed Google Scholar
  5. Martin Srayko
    You can also search for this author inPubMed Google Scholar
  6. Thimo Kurz
    You can also search for this author inPubMed Google Scholar
  7. Sarah Glaser
    You can also search for this author inPubMed Google Scholar
  8. Paul E. Mains
    You can also search for this author inPubMed Google Scholar
  9. Mike Tyers
    You can also search for this author inPubMed Google Scholar
  10. Bruce Bowerman
    You can also search for this author inPubMed Google Scholar
  11. Matthias Peter
    You can also search for this author inPubMed Google Scholar

Corresponding authors

Correspondence toLionel Pintard or Matthias Peter.

Ethics declarations

Competing interests

The authors declare that they have no competing financial interests.

Supplementary information

Rights and permissions

About this article

Cite this article

Pintard, L., Willis, J., Willems, A. et al. The BTB protein MEL-26 is a substrate-specific adaptor of the CUL-3 ubiquitin-ligase.Nature 425, 311–316 (2003). https://doi.org/10.1038/nature01959

Download citation

This article is cited by