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.

  • Short Communication
  • Published:

Disruption of the Pdcd4 tumor suppressor gene in chicken DT40 cells reveals its role in the DNA-damage response

Oncogene volume 28, pages 3758–3764 (2009)Cite this article

Abstract

The programmed cell death gene 4 (Pdcd4) gene has been implicated as a new tumor suppressor gene in the development of several types of human cancer. Pdcd4 interacts with the translation initiation factor, eIF4A, and is thought to act as a translation inhibitor. Here, we have used the chicken B-cell line DT40 to disrupt the Pdcd4 gene by homologous recombination. Our study shows that cells lacking a functional Pdcd4 gene are viable and have no obvious defects when cultivated under normal growth conditions. However, Pdcd4 knockout cells show an increased sensitivity to agents that cause DNA damage, such as UV light, etoposide or ethyl-methanesulfonate. In summary, our findings show that Pdcd4 has an important function in the cellular response to DNA damage. Low Pdcd4 expression, which is frequently observed in tumor cells, might therefore contribute to tumorigenesis by disturbing the cellular DNA-damage response.

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

Similar content being viewed by others

References

  • Asangani IA, Rasheed SA, Nikolova DA, Leupold JH, Colburn NH, Post S et al. (2008). MicroRNA-21 (miR-21) post-transcriptionally downregulates tumor suppressor Pdcd4 and stimulates invasion, intravasation and metastasis in colorectal cancer. Oncogene 27: 2128–2136.

    Article  CAS  Google Scholar 

  • Bezzubova O, Silbergleit A, Yamaguchi-Iwai Y, Takeda S, Buerstedde JM . (1997). Reduced X-ray resistance and homologous recombination frequencies in a RAD54−/− mutant of the chicken DT40 cell line. Cell 89: 185–193.

    Article  CAS  Google Scholar 

  • Bitomsky N, Böhm M, Klempnauer K-H . (2004). Transformation suppressor protein Pdcd4 interferes with JNK-mediated phosphorylation of c-Jun and recruitment of the coactivator p300 by c-Jun. Oncogene 23: 7484–7493.

    Article  CAS  Google Scholar 

  • Bitomsky N, Wethkamp N, Marikkannu R, Klempnauer K-H . (2008). siRNA-mediated knock-down of Pdcd4 expression causes up-regulation of p21(Waf1/Cip1) expression. Oncogene 27: 4820–4829.

    Article  CAS  Google Scholar 

  • Böhm M, Sawicka K, Siebrasse JP, Brehmer-Fastnacht A, Peters R, Klempnauer K-H . (2003). The transformation suppressor protein Pdcd4 shuttles between nucleus and cytoplasm and binds RNA. Oncogene 22: 4905–4910.

    Article  Google Scholar 

  • Buerstedde JM, Takeda S . (1991). Increased ratio of targeted to random integration after transfection of chicken B cell lines. Cell 67: 179–188.

    Article  CAS  Google Scholar 

  • Chang JH, Cho YH, Sohn SY, Choi JM, Kim A, Kim YC et al. (2009). Cyrstal structure of the eIF4A–PDCD4 complex. Proc Natl Acad Sci USA 106: 3148–3153.

    Article  CAS  Google Scholar 

  • Chen Y, Knosel T, Kristiansen G, Pietas A, Garber ME, Matsuhashi S et al. (2003). Loss of PDCD4 expression in human lung cancer correlates with tumour progression and prognosis. J Pathol 200: 640–646.

    Article  CAS  Google Scholar 

  • Cmarik JL, Min H, Hegamyer G, Zhan S, Kulesz-Martin M, Yoshinaga H et al. (1999). Differentially expressed protein Pdcd4 inhibits tumor promoter-induced neoplastic transformation. Proc Natl Acad Sci USA 96: 14037–14042.

    Article  CAS  Google Scholar 

  • Dorello NV, Peschiaroli A, Guardavaccaro D, Colburn NH, Sherman NE, Pagano M . (2006). S6K1- and betaTRCP-mediated degradation of PDCD4 promotes protein translation and cell growth. Science 314: 467–471.

    Article  Google Scholar 

  • Frankel LB, Christoffersen NR, Jacobsen A, Lindow M, Krogh A, Lund AH . (2008). Programmed cell death 4 (PDCD4) is an important functional target of the microRNA miR-21 in breast cancer cells. J Biol Chem 283: 1026–1033.

    Article  CAS  Google Scholar 

  • Göke A, Göke R, Knolle A, Trusheim H, Schmidt H, Wilmen A et al. (2002). DUG is a novel homologue of translation initiation factor 4G that binds eIF4A. Biochem Biophys Res Commun 297: 78–82.

    Article  Google Scholar 

  • Gorospe M, Cirielli C, Wang X, Seth P, Capogrossi MC, Holbrook NJ . (1997). p21(Waf1/Cip1) protects against p53-mediated apoptosis of human melanoma cells. Oncogene 14: 929–935.

    Article  CAS  Google Scholar 

  • Hershey JWB, Merrick WC . (2000). Translational control of gene expression: pathway and mechanism of initiation of protein synthesis. In: Sonenberg N, Hershey JWB, Mathews MB (eds). Cold Spring Harbor Laboratory Press, Cold Spring Harbor: New York. pp 33–88.

  • Jansen AP, Camalier CE, Colburn NH . (2005). Epidermal expression of the translation inhibitor programmed cell death 4 suppresses tumorigenesis. Cancer Res 65: 6034–6041.

    Article  CAS  Google Scholar 

  • LaRonde-Blanc N, Santhanam AN, Baker AR, Wlodawer A, Colburn NH . (2007). Structural basis for inhibition of translation by the tumor suppressor Pdcd4. Mol Cell Biol 27: 147–156.

    Article  Google Scholar 

  • Leupold JH, Yang HS, Colburn NH, Asangani I, Post S, Allgayer H . (2007). Tumor suppressor Pdcd4 inhibits invasion/intravasation and regulates urokinase receptor (u-PAR) gene expression via Sp-transcription factors. Oncogene 26: 4550–4562.

    Article  CAS  Google Scholar 

  • Loh PG, Yang HS, Walsh MA, Wang Q, Wang X, Cheng Z et al. (2009). Structural basis for translational inhibition by the tumour suppressor Pdcd4. EMBO J 28: 274–285.

    Article  CAS  Google Scholar 

  • Lu Z, Liu M, Stribinskis V, Klinge CM, Ramos KS, Colburn NH . (2008). MicroRNA-21 promotes cell transformation by targeting the programmed cell death 4 gene. Oncogene 27: 4373–4379.

    Article  CAS  Google Scholar 

  • Mudduluru G, Medved F, Grobholz R, Jost C, Gruber A, Leupold JH et al. (2007). Loss of programmed cell death 4 expression marks adenoma-carcinoma transition, correlates inversely with phosphorylated protein kinase B, and is an independent prognostic factor in resected colorectal cancer. Cancer 110: 1697–1707.

    Article  CAS  Google Scholar 

  • Palamarchuk A, Efanov A, Maximov V, Aqeilan RI, Croce CM, Pekarsky Y . (2005). Akt phosphorylates and regulates Pdcd4 tumor suppressor protein. Cancer Res 65: 11282–11286.

    Article  CAS  Google Scholar 

  • Schlichter U, Kattmann D, Appl H, Miethe J, Brehmer-Fastnacht A, Klempnauer K-H . (2001). Identification of the myb-inducible promoter of the chicken Pdcd4 gene. Biochim Biophys Acta 1520: 99–104.

    Article  CAS  Google Scholar 

  • Seoane J, Le HV, Massague J . (2002). Myc suppression of the p21(Cip1) Cdk inhibitor influences the outcome of the p53 response to DNA damage. Nature 419: 729–734.

    Article  CAS  Google Scholar 

  • Shibahara K, Asano M, Ishida Y, Aoki T, Koike T, Honjo T . (1995). Isolation of a novel mouse gene MA-3 that is induced upon programmed cell death. Gene 166: 297–301.

    Article  CAS  Google Scholar 

  • Suzuki C, Garces RG, Edmonds KA, Hiller S, Hyberts SG, Marintchev A et al. (2008). PDCD4 inhibits translation initiation by binding to eIF4A using both its MA3 domains. Proc Natl Acad Sci USA 105: 3274–3279.

    Article  CAS  Google Scholar 

  • Takao N, Kato H, Mori R, Morrison C, Sonada E, Sun X et al. (1999). Disruption of ATM in p53-null cells causes multiple functional abnormalities in cellular response to ionizing radiation. Oncogene 18: 7002–7009.

    Article  CAS  Google Scholar 

  • Wang Q, Sun Z, Yang H-S . (2008). Downregulation of tumour suppressor Pdcd4 promotes invasion and activates both β-catenin/TCF and AP-1-dependent transcription in colon carcinoma cells. Oncogene 27: 1527–1535.

    Article  CAS  Google Scholar 

  • Waters LC, Veverka V, Böhm M, Schmedt T, Choong PT, Muskett FW et al. (2007). Structure of the C-terminal MA-3 domain of the tumour suppressor protein Pdcd4 and characterization of its interaction with eIF4A. Oncogene 26: 4941–4950.

    Article  CAS  Google Scholar 

  • Yang HS, Cho MH, Zacowicz H, Hegamyer G, Sonenberg N, Colburn N . (2004). A novel function of the MA-3 domains in transformation and translation suppressor Pdcd4 is essential for its binding to eukaryotic translation initiation factor 4A. Mol Cell Biol 24: 3894–3906.

    Article  CAS  Google Scholar 

  • Yang HS, Jansen AP, Komar AA, Zheng X, Merrick WC, Costes S et al. (2003a). The transformation suppressor Pdcd4 is a novel eukaryotic translation initiation factor 4A binding protein that inhibits translation. Mol Cell Biol 23: 26–37.

    Article  Google Scholar 

  • Yang HS, Knies JL, Stark C, Colburn NH . (2003b). Pdcd4 suppresses tumor phenotype in JB6 cells by inhibiting AP-1 transactivation. Oncogene 22: 3712–3720.

    Article  CAS  Google Scholar 

  • Yang HS, Matthews CP, Clair T, Wang Q, Baker AR, Li C-CH et al. (2006). Tumorigenesis suppressor Pdcd4 down-regulates mitogen-activated protein kinase kinase kinase kinase 1 expression to suppress colon carcinoma cell invasion. Mol Cell Biol 26: 1297–1306.

    Article  Google Scholar 

  • Zhang H, Ozaki I, Mizuta T, Hamajima H, Yasutake T, Eguchi Y et al. (2006). Involvement of programmed cell death 4 in transforming growth factor-beta1-induced apoptosis in human hepatocellular carcinoma. Oncogene 25: 6101–6112.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank B Berkenfeld for excellent technical assistance. This study was supported by grants from the Deutsche Krebshilfe (10-1716) and the Wilhelm-Sander-Stiftung (2004.088.1). PS and RM were supported by fellowships from the Graduate School of Chemistry (GSC-MS) at the University of Münster.

Author information

Author notes

  1. N Bitomsky

    Present address: 2Current address: German Cancer Research Center, Im Neuenheimer Feld 280, Heidelberg D-69120, Germany,

Authors and Affiliations

  1. Institute for Biochemistry, Westfälische-Wilhelms-Universität Münster, Münster, Germany

    P Singh, R Marikkannu & K -H Klempnauer

Authors
  1. P Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. R Marikkannu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. N Bitomsky
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. K -H Klempnauer
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to K -H Klempnauer.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Singh, P., Marikkannu, R., Bitomsky, N. et al. Disruption of the Pdcd4 tumor suppressor gene in chicken DT40 cells reveals its role in the DNA-damage response. Oncogene 28, 3758–3764 (2009). https://doi.org/10.1038/onc.2009.239

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2009.239

Keywords

This article is cited by

Search

Advanced search

Quick links