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:

Growth inhibition by CDK-cyclin and PCNA binding domains of p21 occurs by distinct mechanisms and is regulated by ubiquitin-proteasome pathway

Oncogene volume 18, pages 3290–3302 (1999)Cite this article

Abstract

The CDK inhibitor, p21WAF1/Cip1 blocks cell cycle progression. In vitro, the N-terminus of p21 binds and inhibits CDK-cyclin kinase activity, whereas the C-terminus binds and inhibits PCNA (proliferating cell nuclear antigen) function. PCNA is essential for processivity of both DNA polymerase δ and ε. We have performed a detailed analysis of growth inhibition by the N- and C-terminal regions of p21, and determined whether the N- and C-terminal regions mediate this effect by different mechanisms. Expression of either the N- or the C-terminal region of p21 inhibits DNA synthesis and cell growth, but not as efficiently as full length p21. The effectiveness of the two p21 domains is dependent on their stability which is determined by the ubiquitin-proteasome pathway. The stabilization of the N- and C-terminal region of p21 increases their effectiveness as inhibitors of DNA synthesis to levels comparable to full length p21. Inhibition of DNA synthesis by the N-terminal region of p21 involves suppression of E2F activity. In contrast, inhibition by the C-terminal region of p21 is not accompanied by suppression of E2F activity, but is mediated via PCNA binding. The C-terminal region of p21 therefore inhibits cell growth by a mechanism distinct from that of the N-terminal region containing the CDK-cyclin inhibitory domain.

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
Figure 7

Similar content being viewed by others

References

  • Adams PD, Sellers WR, Sharma SK, Wu AD, Nalin CM and Kaelin Jr WG. . 1996 Mol. Cell. Biol. 16: 6623–6633.

  • Ball KL, Lain S, Fahraeus R, Smythe C and Lane DP. . 1997 Curr. Biol. 7: 71–80.

  • Brenot-Bosc F, Gupta S, Margolis RL and Fotedar R. . 1995 Chromosoma 103: 517–527.

  • Cayrol C, Knibiehler M and Ducommun B. . 1998 Oncogene 16: 311–320.

  • Chen IT, Akamatsu M, Smith ML, Lung FD, Duba D, Roller PP, Fornace Jr AJ and O'Connor PM. . 1996 Oncogene 12: 595–607.

  • Chen J, Jackson PK, Kirschner MW and Dutta A. . 1995 Nature 374: 386–388.

  • Chowdary DR, Dermody JJ, Jha KK and Ozer HL. . 1994 Mol. Cell. Biol. 14: 1997–2003.

  • Dimri GP, Nakanishi M, Desprez PY, Smith JR and Campisi J. . 1996 Mol. Cell. Biol. 16: 2987–2997.

  • Dingwall C and Laskey RA. . 1991 Trends Biochem Sci. 16: 478–481.

  • el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, Lin D, Mercer WE, Kinzler KW and Vogelstein B. . 1993 Cell 75: 817–825.

  • Elledge SJ and Harper JW. . 1994 Curr. Opin. Cell. Biol. 6: 847–852.

  • Fenteany G, Standaert RF, Lane WS, Choi S, Corey EJ and Schreiber SL. . 1995 Science 268: 726–731.

  • Flores-Rozas H, Kelman Z, Dean FB, Pan ZQ, Harper JW, Elledge SJ, O'Donnell M and Hurwitz J. . 1994 Proc. Natl. Acad. Sci. USA 91: 8655–8659.

  • Fotedar A, Cannella D, Fitzgerald P, Rousselle T, Gupta S, Doree M and Fotedar R. . 1996a J. Biol. Chem. 271: 31627–31637.

  • Fotedar R, Fitzgerald P, Rousselle T, Cannella D, Doree M, Messier H and Fotedar A. . 1996b Oncogene 12: 2155–2164.

  • Fotedar R, Flatt J, Gupta S, Margolis RL, Fitzgerald P, Messier H and Fotedar A. . 1995 Mol. Cell. Biol. 15: 932–942.

  • Fotedar R, Mossi R, Fitzgerald P, Rousselle T, Maga G, Brickner H, Messier H, Khastilba S, Hubscher U and Fotedar A. . 1996c EMBO J. 15: 4423–4433.

  • Fotedar R and Roberts JM. . 1991 Cold Spring Harb. Symp. Quant. Biol. 56: 325–333.

  • Girard F, Strausfeld U, Fernandez A and Lamb NJ. . 1991 Cell 67: 1169–1179.

  • Goubin F and Ducommun B. . 1995 Oncogene 10: 2281–2287.

  • Gu Y, Rosenblatt J and Morgan DO. . 1992 EMBO J. 11: 3995–4005.

  • Harper JW, Adami GR, Wei N, Keyomarsi K and Elledge SJ. . 1993 Cell 75: 805–816.

  • Harper JW, Elledge SJ, Keyomarsi K, Dynlacht B, Tsai LH, Zhang P, Dobrowolski S, Bai C, Connell-Crowley L, Swindell E, Fox MP and Wei N. . 1995 Mol. Biol. Cell. 6: 387–400.

  • Hurwitz J, Dean FB, Kwong AD and Lee SH. . 1990 J. Biol. Chem. 265: 18043–18046.

  • Lee MH, Reynisdottir I and Massague J. . 1995 Genes Dev. 9: 639–649.

  • Li R, Hannon GJ, Beach D and Stillman B. . 1996 Curr. Biol. 6: 189–199.

  • Li R, Waga S, Hannon GJ, Beach D and Stillman B. . 1994 Nature 371: 534–537.

  • Lin J, Reichner C, Wu X and Levine AJ. . 1996 Mol. Cell. Biol. 16: 1786–1793.

  • Lukas J, Petersen BO, Holm K, Bartek J and Helin K. . 1996 Mol. Cell. Biol. 16: 1047–1057.

  • Luo Y, Hurwitz J and Massague J. . 1995 Nature 375: 355–363.

  • Maki CG and Howley PM. . 1997 Mol. Cell. Biol. 17: 355–363.

  • Matsuoka S, Edwards MC, Bai C, Parker S, Zhang P, Baldini A, Harper JW and Elledge SJ. . 1995 Genes Dev. 9: 650–662.

  • Nakanishi M, Robetorye RS, Pereira-Smith OM and Smith JR. . 1995 J. Biol. Chem. 270: 17060–17063.

  • Nigg EA. . 1993 Curr. Opin. Cell. Biol. 5: 187–193.

  • Nigg EA. . 1997 Nature 386: 779–787.

  • Noda A, Ning Y, Venable SF, Pereira-Smith OM and Smith JR. . 1994 Exp. Cell. Res. 211: 90–98.

  • Ogryzko VV, Wong P and Howard BH. . 1997 Mol. Cell. Biol. 17: 4877–4882.

  • Pan ZQ, Reardon JT, Li L, Flores-Rozas H, Legerski R, Sancar A and Hurwitz J. . 1995 J. Biol. Chem. 270: 22008–22016.

  • Polyak K, Lee MH, Erdjument-Bromage H, Koff A, Roberts JM, Tempst P and Massague J. . 1994 Cell 78: 59–66.

  • Roberts JM, Koff A, Polyak K, Firpo E, Collins S, Ohtsubo M and Massague J. . 1994 Cold Spring Harb. Symp. Quant. Biol. 59: 31–38.

  • Russo AA, Jeffrey PD and Pavletich NP. . 1996 Nat. Struct. Biol. 3: 696–700.

  • Sasaki T, Kishi M, Saito M, Tanaka T, Higuchi N, Kominam E, Katunuma N and Murachi T. . 1990 J. Enzyme Inhib. 3: 195–201.

  • Sherr CJ and Roberts JM. . 1995 Genes Dev. 9: 1149–1163.

  • Shivji KK, Kenny MK and Wood RD. . 1992 Cell 69: 367–374.

  • Shivji MK, Grey SJ, Strausfeld UP, Wood RD and Blow JJ. . 1994 Curr. Biol. 4: 1062–1068.

  • Shivji MK, Podust VN, Hubscher U and Wood RD. . 1995 Biochemistry 34: 5011–5017.

  • Shiyanov P, Bagchi S, Adami G, Kokontis J, Hay N, Arroyo M, Morozov A and Raychaudhuri P. . 1996 Mol. Cell. Biol. 16: 737–744.

  • Stillman B. . 1994 Cell 78: 725–728.

  • Toyoshima H and Hunter T. . 1994 Cell 78: 67–74.

  • Treier M, Staszewski LM and Bohmann D. . 1994 Cell 78: 787–798.

  • Waga S, Hannon GJ, Beach D and Stillman B. . 1994 Nature 369: 574–578.

  • Warbrick E, Lane DP, Glover DM and Cox LS. . 1995 Curr. Biol. 5: 275–282.

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

  • Xiong Y, Hannon GJ, Zhang H, Casso D, Kobayashi R and Beach D. . 1993a Nature 366: 701–704.

  • Xiong Y, Zhang H and Beach D. . 1993b Genes Dev. 7: 1572–1583.

  • Zhu L, Harlow E and Dynlacht BD. . 1995 Genes Dev. 9: 1740–1752.

  • Zhu L, van den Heuvel S, Helin K, Fattaey A, Ewen M, Livingston D, Dyson N and Harlow E. . 1993 Genes Dev. 7: 1111–1125.

Download references

Acknowledgements

The authors acknowledge the kind gifts of plasmids and reagents by K Helin (European Institute of Oncology, Milan, Spain), N Dyson and E Harlow (Harvard Medical School, Boston, MA, USA), D Bohmann and L Staszewski (EMBL, Heidelberg, Germany). We thank Howard Brickner for technical assistance. Jerome Boulaire is a recipient of doctoral fellowship from Region Rhône Alpes. Denis Rousseau was a recipient of fellowship from Société de Secours des Amis des Sciences. This work was supported by grants from INSERM (#930104), ARC (#9621) and Region Rhone Alpes (HO9873.08.12) to Rati Fotedar and by grants from the NIH (#AI31453, #CA74435) to Arun Fotedar.

Author information

Author notes

  1. Rati Fotedar: Author for correspondence

  2. Denis Rousseau and Dominique Cannella: D Rousseau and D Cannella contributed equally to the paper

Authors and Affiliations

  1. Institut de Biologie Structurale J-P Ebel, 41 avenue des Martyrs, 38027, Grenoble, Cedex 1, France

    Denis Rousseau, Dominique Cannella, Jerome Boulaire & Rati Fotedar

  2. Sidney Kimmel Cancer Center, 10835 Altman Road, San Diego, California, CA 92121, USA

    Patrick Fitzgerald & Arun Fotedar

Authors
  1. Denis Rousseau
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dominique Cannella
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jerome Boulaire
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Patrick Fitzgerald
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Arun Fotedar
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Rati Fotedar
    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

Rousseau, D., Cannella, D., Boulaire, J. et al. Growth inhibition by CDK-cyclin and PCNA binding domains of p21 occurs by distinct mechanisms and is regulated by ubiquitin-proteasome pathway. Oncogene 18, 3290–3302 (1999). https://doi.org/10.1038/sj.onc.1202681

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1202681

Keywords

Search

Advanced search

Quick links