Cytoplasmic ubiquitin ligase KPC regulates proteolysis of p27Kip1 at G1 phase (original) (raw)

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

Abstract

The cyclin-dependent kinase inhibitor p27_Kip1_ 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 p27_Kip1_ degradation3,4,5,6, proteolysis of p27_Kip1_ at the G0–G1 transition proceeds normally in Skp2 −/− cells7,8. Moreover, p27_Kip1_ 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 p27_Kip1_ 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 p27_Kip1_ and is localized to the cytoplasm. Overexpression of KPC promoted the degradation of p27_Kip1_, whereas a dominant-negative mutant of KPC1 delayed p27_Kip1_ degradation. The nuclear export of p27_Kip1_ by CRM1 seems to be necessary for KPC-mediated proteolysis. Depletion of KPC1 by RNA interference also inhibited p27_Kip1_ degradation. KPC thus probably controls degradation of p27_Kip1_ in G1 phase after export of the latter from the nucleus.

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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.

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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
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  2. Taichi Hara
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  3. Masaki Matsumoto
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  4. Noriko Ishida
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  5. Fumihiko Okumura
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  6. Shigetsugu Hatakeyama
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  7. Minoru Yoshida
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  8. Keiko Nakayama
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  9. Keiichi I. Nakayama
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Correspondence toKeiichi I. Nakayama.

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Kamura, T., Hara, T., Matsumoto, M. et al. Cytoplasmic ubiquitin ligase KPC regulates proteolysis of p27_Kip1_ at G1 phase.Nat Cell Biol 6, 1229–1235 (2004). https://doi.org/10.1038/ncb1194

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