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.

  • Original Paper
  • Published:

PKCη associates with cyclin E/cdk2/p21 complex, phosphorylates p21 and inhibits cdk2 kinase in keratinocytes

Oncogene volume 19, pages 6334–6341 (2000)Cite this article

Abstract

PKC is activated on the cell membrane by phospholipids, thereby transducing signals to intracellular pathways. We provide here another function of PKC, namely, regulating cell cycle by interaction with the cyclin E/cdk2/p21 complex. Among the 10 isoforms of PKC, PKCη is predominantly expressed in squamous cell epithelia and induces terminal differentiation of keratinocytes. PKCη that is endogenously expressed or overexpressed was found to associate with the cyclin E/cdk2/p21 complex in keratinocytes of mice and humans. Requirement of a possible adaptor protein to the binding was suggested by the reconstitution of PKCη and the cyclin E/cdk2/p21 complex which were prepared from human keratinocytes or Sf9 insect cells. Colocalization of PKCη with cdk2 and cyclin E was observed in the cytoplasm, particularly in the perinuclear region. p21 was phosphorylated in the complex in a PKC-activator dependent manner. Association of PKCη with cdk2 resulted in marked inhibition of cdk2-kinase activity when measured by phosphorylation of Rb. Dominant negative PKCη associated with the cyclin E/cdk2/p21 complex, but caused a little inhibition of cdk2 kinase activity. Among the known regulatory mechanisms of cdk2 activity, dephosphorylation of Thr160 was demonstrated.

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
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  • Ashton AW, Watanabe G, Albanese C, Harrington EO, Ware JA and Pestell RG. . 1999 J. Biol. Chem. 274: 20805–20811.

  • Chapline C, Cottom J, Tobin H, Hulmes J, Crabb J and Jaken S. . 1998 J. Biol. Chem. 273: 19482–19489.

  • Chida K, Sagara H, Suzuki Y, Murakami A, Osada S, Ohno S, Hirosawa K and Kuroki T. . 1994 Mol. Cell Biol. 14: 3782–3790.

  • Di Cunto F, Topley G, Calautti E, Hsiao J, Ong L, Seth PK and Dotto GP. . 1998 Science 280: 1069–1072.

  • Diaz-Meco MT, Municio MM, Frutos S, Sanchez P, Lozano J, Sanz L and Moscat J. . 1996 Cell 86: 777–786.

  • Doornbos RP, Theelen M, van der Hoeven PC, van Blitterswijk WJ, Verkleij AJ and van Bergen en Henegouwen PM. . 1999 J. Biol. Chem. 274: 8589–8596.

  • Elledge SJ. . 1996 Science 274: 1664–1672.

  • Fisher RP, Jin P, Chamberlin HM and Morgan DO. . 1995 Cell 83: 47–57.

  • Fisher RP and Morgan DO. . 1994 Cell 78: 713–724.

  • Fujise A, Mizuno K, Ueda Y, Osada S, Hirai S, Takayanagi A, Shimizu N, Owada MK, Nakajima H and Ohno S. . 1994 J. Biol. Chem. 269: 31642–31648.

  • Harrington EO, Loffler J, Nelson PR, Kent KC, Simons M and Ware JA. . 1997 J. Biol. Chem. 272: 7390–7397.

  • Ikuta T, Chida K, Tajima O, Matsuura Y, Iwamori M, Ueda Y, Mizuno K, Ohno S and Kuroki T. . 1994 Cell Growth Differ. 5: 943–947.

  • Ishino K, Ohba M, Kashiwagi M, Kawabe S, Chida K and Kuroki T. . 1998 Jpn. J. Cancer Res. 89: 1126–1133.

  • Izumi Y, Hirose T, Tamai Y, Hirai S, Nagashima Y, Fujimoto T, Tabuse Y, Kemphues KJ and Ohno S. . 1998 J. Cell Biol. 143: 95–106.

  • Jain N, Zhang T, Kee WH, Li W and Cao X. . 1999 J. Biol. Chem. 274: 24392–24400.

  • Jaumot M, Estanol JM, Casanovas O, Grana X, Agell N and Bachs O. . 1997 Biochem. Biophys. Res. Commun. 241: 434–438.

  • Kanegae Y, Makimura M and Saito I. . 1994 Jpn. J. Med. Sci. Biol. 47: 157–166.

  • Konishi H, Kuroda S and Kikkawa U. . 1994 Biochem. Biophys. Res. Commun. 205: 1770–1775.

  • Kuroki T, Kashiwagi M, Ishino K, Huh N and Ohba M. . 1999 J. Investig. Dermatol. Symp. Proc. 4: 153–157.

  • Le Good JA, Ziegler WH, Parekh DB, Alessi DR, Cohen P and Parker PJ. . 1998 Science 281: 2042–2045.

  • Livneh E, Shimon T, Bechor E, Doki Y, Schieren I and Weinstein IB. . 1996 Oncogene 12: 1545–1555.

  • Missero C, Di Cunto F, Kiyokawa H, Koff A and Dotto GP. . 1996 Genes Dev. 10: 3065–3075.

  • Miyake S, Makimura M, Kanegae Y, Harada S, Sato Y, Takamori K, Tokuda C and Saito I. . 1996 Proc. Natl. Acad. Sci. USA 93: 1320–1324.

  • Mochly-Rosen D and Gordon AS. . 1998 FASEB J. 12: 35–42.

  • Morgan DO. . 1995 Nature 374: 131–134.

  • Morgan DO. . 1997 Annu. Rev. Cell Dev. Biol. 13: 261–291.

  • Morrow TA, Muljo SA, Zhang J, Hardwick JM and Schlissel MS. . 1999 Mol. Cell. Biol. 19: 5608–5618.

  • Nishizuka Y. . 1988 Nature 334: 661–665.

  • Nishizuka Y. . 1992 Science 258: 607–614.

  • Ohba M, Ishino K, Kashiwagi M, Kawabe S, Chida K, Huh NH and Kuroki T. . 1998 Mol. Cell Biol. 18: 5199–5207.

  • Osada S, Hashimoto Y, Nomura S, Kohno Y, Chida K, Tajima O, Kubo K, Akimoto K, Koizumi H, Kitamura Y and et al. 1993 Cell Growth Differ. 4: 167–175.

  • Osada S, Mizuno K, Saido TC, Akita Y, Suzuki K, Kuroki T and Ohno S. . 1990 J. Biol. Chem. 265: 22434–22440.

  • Pal S, Claffey KP, Dvorak HF and Mukhopadhyay D. . 1997 J. Biol. Chem. 272: 27509–27512.

  • Poon RY and Hunter T. . 1995 Science 270: 90–93.

  • Puls A, Schmidt S, Grawe F and Stabel S. . 1997 Proc. Natl. Acad. Sci. USA 94: 6191–6196.

  • Resnick MS, Kang BS, Luu D, Wickham JT, Sando JJ and Hahn CS. . 1998 J. Biol. Chem. 273: 27654–27661.

  • Romanelli A, Martin KA, Toker A and Blenis J. . 1999 Mol. Cell Biol. 19: 2921–2928.

  • Ron D, Napolitano EW, Voronova A, Vasquez NJ, Roberts DN, Calio BL, Caothien RH, Pettiford SM, Wellik S, Mandac JB and Kauvar LM. . 1999 J. Biol. Chem. 274: 19003–19010.

  • Sanz L, Sanchez P, Lallena MJ, Diaz-Meco MT and Moscat J. . 1999 EMBO J. 18: 3044–3053.

  • Scott MT, Morrice N and Ball KL. . 2000 J. Biol. Chem. 275: 11529–11537.

  • Sherr CJ. . 1996 Science 274: 1672–1677.

  • Slosberg ED, Klein MG, Yao Y, Han EK, Schieren I and Weinstein IB. . 1999 Oncogene 18: 6658–6666.

  • Staudinger J, Lu J and Olson EN. . 1997 J. Biol. Chem. 272: 32019–32024.

  • Ueffing M, Lovric J, Philipp A, Mischak H and Kolch W. . 1997 Oncogene 15: 2921–2927.

  • Waldron RT, Iglesias T and Rozengurt E. . 1999 J. Biol. Chem. 274: 9224–9230.

  • Watanabe H, Pan ZQ, Schreiber-Agus N, DePinho RA, Hurwitz J and Xiong Y. . 1998 Proc. Natl. Acad. Sci. USA 95: 1392–1397.

  • Watanabe T, Ono Y, Taniyama Y, Hazama K, Igarashi K, Ogita K, Kikkawa U and Nishizuka Y. . 1992 Proc. Natl. Acad. Sci. USA 89: 10159–10163.

  • Weinberg RA. . 1995 Cell 81: 323–330.

  • Yao L, Kawakami Y and Kawakami T. . 1994 Proc. Natl. Acad. Sci. USA 91: 9175–9179.

  • Yaroslavskiy B, Watkins S, Donnenberg AD, Patton TJ and Steinman RA. . 1999 Blood 93: 2907–2917.

  • Zhang P, Wong C, Liu D, Finegold M, Harper JW and Elledge SJ. . 1999 Genes Dev. 13: 213–224.

Download references

Acknowledgements

The authors acknowledge the kind help of adenovirus vector construction and kind gift of Ax-lacZ vector by Yumi Kanegae and Izumu Saito (Institute of Medical Science, University of Tokyo). This work was supported in part by a Grant-in-aid for Cancer Research from the Ministry of Education, Science, Sports and Culture of Japan.

Author information

Authors and Affiliations

  1. Institute of Molecular Oncology, Showa University, 1-5-8 Hatanodai, Shinagawa-ku, Tokyo, 142-8555, Japan

    Mariko Kashiwagi, Motoi Ohba, Hiroyuki Watanabe, Keiko Ishino & Toshio Kuroki

  2. Department of Biochemical Cell Research, The Tokyo Metropolitan Institute of Medical Science, 3-18-22 Honkomagome, Bunkyo-ku, Tokyo, 113-0021, Japan

    Kohji Kasahara & Yutaka Sanai

  3. Biology Division, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo, 104-0045, Japan

    Yoichi Taya

Authors
  1. Mariko Kashiwagi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Motoi Ohba
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hiroyuki Watanabe
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Keiko Ishino
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kohji Kasahara
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yutaka Sanai
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yoichi Taya
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Toshio Kuroki
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kashiwagi, M., Ohba, M., Watanabe, H. et al. PKCη associates with cyclin E/cdk2/p21 complex, phosphorylates p21 and inhibits cdk2 kinase in keratinocytes. Oncogene 19, 6334–6341 (2000). https://doi.org/10.1038/sj.onc.1204028

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1204028

Keywords

This article is cited by

Search

Advanced search

Quick links