Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Cytoplasmic ubiquitin ligase KPC regulates proteolysis of p27Kip1 at G1 phase

Nature Cell Biology volume 6, pages 1229–1235 (2004)Cite this article

Abstract

The cyclin-dependent kinase inhibitor p27Kip1 is degraded at the G0–G1 transition of the cell cycle by the ubiquitin–proteasome pathway1,2. Although the nuclear ubiquitin ligase (E3) SCFSkp2 is implicated in p27Kip1 degradation3,4,5,6, proteolysis of p27Kip1 at the G0–G1 transition proceeds normally in Skp2−/− cells7,8. Moreover, p27Kip1 is exported from the nucleus to the cytoplasm at G0–G1 (refs 9–11). These data suggest the existence of a Skp2-independent pathway for the degradation of p27Kip1 at G1 phase. We now describe a previously unidentified E3 complex: KPC (Kip1 ubiquitination-promoting complex), consisting of KPC1 and KPC2. KPC1 contains a RING-finger domain, and KPC2 contains a ubiquitin-like domain and two ubiquitin-associated domains. KPC interacts with and ubiquitinates p27Kip1 and is localized to the cytoplasm. Overexpression of KPC promoted the degradation of p27Kip1, whereas a dominant-negative mutant of KPC1 delayed p27Kip1 degradation. The nuclear export of p27Kip1 by CRM1 seems to be necessary for KPC-mediated proteolysis. Depletion of KPC1 by RNA interference also inhibited p27Kip1 degradation. KPC thus probably controls degradation of p27Kip1 in G1 phase after export of the latter from the nucleus.

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

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

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

Figure 1: Purification of KPC.
Figure 2: Polyubiquitination of p27Kip1 by KPC.
Figure 3: Interaction between KPC and p27Kip1 and subcellular localization of KPC.
Figure 4: Effect of KPC overexpression or depletion on p27Kip1 degradation.
Figure 5: Effects of KPC1 depletion on p27Kip1 degradation and cell-cycle progression.

Similar content being viewed by others

Accession codes

Accessions

BINDPlus

GenBank/EMBL/DDBJ

References

  1. Pagano, M. et al. Role of the ubiquitin–proteasome pathway in regulating abundance of the cyclin-dependent kinase inhibitor p27. Science 269, 682–685 (1995).

    Article  CAS  Google Scholar 

  2. Shirane, M. et al. Down-regulation of p27Kip1 by two mechanisms, ubiquitin-mediated degradation and proteolytic processing. J. Biol. Chem. 274, 13886–13893 (1999).

    Article  CAS  Google Scholar 

  3. Carrano, A. C., Eytan, E., Hershko, A. & Pagano, M. SKP2 is required for ubiquitin-mediated degradation of the CDK inhibitor p27. Nature Cell Biol. 1, 193–199 (1999).

    Article  CAS  Google Scholar 

  4. Sutterluty, H. et al. p45SKP2 promotes p27Kip1 degradation and induces S phase in quiescent cells. Nature Cell Biol. 1, 207–214 (1999).

    Article  CAS  Google Scholar 

  5. Tsvetkov, L. M., Yeh, K. H., Lee, S. J., Sun, H. & Zhang, H. p27Kip1 ubiquitination and degradation is regulated by the SCFSkp2 complex through phosphorylated Thr187 in p27. Curr. Biol. 9, 661–664 (1999).

    Article  CAS  Google Scholar 

  6. Nakayama, K. et al. Targeted disruption of Skp2 results in accumulation of cyclin E and p27Kip1, polyploidy and centrosome overduplication. EMBO J. 19, 2069–2081 (2000).

    Article  CAS  Google Scholar 

  7. Hara, T. et al. Degradation of p27Kip1 at the G0-G1 transition mediated by a Skp2-independent ubiquitination pathway. J. Biol. Chem. 276, 48937–48943 (2001).

    Article  CAS  Google Scholar 

  8. Nakayama, K. et al. Skp2-mediated degradation of p27 regulates progression into mitosis. Dev. Cell 6, 661–672 (2004).

    Article  CAS  Google Scholar 

  9. Ishida, N. et al. Phosphorylation of p27Kip1 on serine 10 is required for its binding to CRM1 and nuclear export. J. Biol. Chem. 277, 14355–14358 (2002).

    Article  CAS  Google Scholar 

  10. Tomoda, K., Kubota, Y. & Kato, J. Degradation of the cyclin-dependent-kinase inhibitor p27Kip1 is instigated by Jab1. Nature 398, 160–165 (1999).

    Article  CAS  Google Scholar 

  11. Rodier, G. et al. p27 cytoplasmic localization is regulated by phosphorylation on Ser10 and is not a prerequisite for its proteolysis. EMBO J. 20, 6672–6682 (2001).

    Article  CAS  Google Scholar 

  12. Li, C., Rodriguez, M., Adamson, J. W. & Banerjee, D. Identification of a glialblastoma cell differentiation factor-related gene mRNA in human microvascular endothelial cells. Genomics 65, 243–252 (2000).

    Article  CAS  Google Scholar 

  13. Ponting, C., Schultz, J. & Bork, P. SPRY domains in ryanodine receptors (Ca2+-release channels). Trends Biochem. Sci. 22, 193–194 (1997).

    Article  CAS  Google Scholar 

  14. Wilkinson, C. R. et al. Proteins containing the UBA domain are able to bind to multi-ubiquitin chains. Nature Cell Biol. 3, 939–943 (2001).

    Article  CAS  Google Scholar 

  15. Chen, L., Shinde, U., Ortolan, T. G. & Madura, K. Ubiquitin-associated (UBA) domains in Rad23 bind ubiquitin and promote inhibition of multi-ubiquitin chain assembly. EMBO Rep. 2, 933–938 (2001).

    Article  CAS  Google Scholar 

  16. Yoshida, M. & Horinouchi, S. Trichostatin and leptomycin. Inhibition of histone deacetylation and signal-dependent nuclear export. Ann. NY Acad. Sci. 886, 23–36 (1999).

    Article  CAS  Google Scholar 

  17. Liang, J. et al. PKB/Akt phosphorylates p27, impairs nuclear import of p27 and opposes p27-mediated G1 arrest. Nature Med. 8, 1153–1160 (2002).

    Article  CAS  Google Scholar 

  18. Viglietto, G. et al. Cytoplasmic relocalization and inhibition of the cyclin-dependent kinase inhibitor p27Kip1 by PKB/Akt-mediated phosphorylation in breast cancer. Nature Med. 8, 1136–1144 (2002).

    Article  CAS  Google Scholar 

  19. Shin, I. et al. PKB/Akt mediates cell-cycle progression by phosphorylation of p27Kip1 at threonine 157 and modulation of its cellular localization. Nature Med. 8, 1145–1152 (2002).

    Article  CAS  Google Scholar 

  20. Bornstein, G. et al. Role of the SCFSkp2 ubiquitin ligase in the degradation of p21Cip1 in S phase. J. Biol. Chem. 278, 25752–25757 (2003).

    Article  CAS  Google Scholar 

  21. Kamura, T. et al. Degradation of p57Kip2 mediated by SCFSkp2-dependent ubiquitination. Proc. Natl Acad. Sci. USA 100, 10231–10236 (2003).

    Article  CAS  Google Scholar 

  22. Hershko, A., Heller, H., Elias, S. & Ciechanover, A. Components of ubiquitin-protein ligase system. Resolution, affinity purification, and role in protein breakdown. J. Biol. Chem. 258, 8206–8214 (1983).

    CAS  PubMed  Google Scholar 

  23. Kamura, T. et al. The Elongin BC complex interacts with the conserved SOCS-box motif present in members of the SOCS, ras, WD-40 repeat, and ankyrin repeat families. Genes Dev. 12, 3872–3881 (1998).

    Article  CAS  Google Scholar 

  24. Kamura, T. et al. Rbx1, a component of the VHL tumor suppressor complex and SCF ubiquitin ligase. Science 284, 657–661 (1999).

    Article  CAS  Google Scholar 

  25. Morita, S., Kojima, T. & Kitamura, T. Plat-E: an efficient and stable system for transient packaging of retroviruses. Gene Ther. 7, 1063–1066 (2000).

    Article  CAS  Google Scholar 

  26. Nakayama, K. et al. Mice lacking p27Kip1 display increased body size, multiple organ hyperplasia, retinal dysplasia, and pituitary tumors. Cell 85, 707–720 (1996).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank T. Kitamura for pMX-puro; R. Yada, N. Nishimura and S. Matsushita for technical assistance; and M. Kimura, A. Ohta and C. Sugita for help in preparation of the manuscript. This work was supported in part by a grant from the Ministry of Education, Science, Sports and Culture of Japan, and by a research grant from the Human Frontier Science Program.

Author information

Authors and Affiliations

  1. Department of Molecular and Cellular Biology, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, Fukuoka, 812-8582, Japan

    Takumi Kamura, Taichi Hara, Masaki Matsumoto, Fumihiko Okumura, Shigetsugu Hatakeyama & Keiichi I. Nakayama

  2. CREST, Japan Science and Technology Corporation, 4-1-8 Honcho, Kawaguchi, Saitama, 332-0012, Japan

    Takumi Kamura, Taichi Hara, Masaki Matsumoto, Noriko Ishida, Fumihiko Okumura, Shigetsugu Hatakeyama, Keiko Nakayama & Keiichi I. Nakayama

  3. Department of Developmental Biology, Center for Translational and Advanced Animal Research on Human Disease, Graduate School of Medicine, Tohoku University, 2-1, Seiryo-machi, Aoba-ku, Sendai, 980-8575, Japan

    Noriko Ishida & Keiko Nakayama

  4. Chemical Genetics Laboratory, Discovery Research Institute, RIKEN, Hirosawa 2-1, Wako, Saitama, 351-0198, Japan

    Minoru Yoshida

Authors
  1. Takumi Kamura
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Taichi Hara
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Masaki Matsumoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Noriko Ishida
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Fumihiko Okumura
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shigetsugu Hatakeyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Minoru Yoshida
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Keiko Nakayama
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Keiichi I. Nakayama
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Keiichi I. Nakayama.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Fig. S1, S2, S3 and S4 (PDF 278 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kamura, T., Hara, T., Matsumoto, M. et al. Cytoplasmic ubiquitin ligase KPC regulates proteolysis of p27Kip1 at G1 phase. Nat Cell Biol 6, 1229–1235 (2004). https://doi.org/10.1038/ncb1194

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb1194

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing