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.

  • Oncogenomics
  • Published:

Transcriptomal profiling of the cellular transformation induced by Rho subfamily GTPases

Oncogene volume 26, pages 4295–4305 (2007)Cite this article

Abstract

We have used microarray technology to identify the transcriptional targets of Rho subfamily guanosine 5′-triphosphate (GTP)ases in NIH3T3 cells. This analysis indicated that murine fibroblasts transformed by these proteins show similar transcriptomal profiles. Functional annotation of the regulated genes indicate that Rho subfamily GTPases target a wide spectrum of functions, although loci encoding proteins linked to proliferation and DNA synthesis/transcription are upregulated preferentially. Rho proteins promote four main networks of interacting proteins nucleated around E2F, c-Jun, c-Myc and p53. Of those, E2F, c-Jun and c-Myc are essential for the maintenance of cell transformation. Inhibition of Rock, one of the main Rho GTPase targets, leads to small changes in the transcriptome of Rho-transformed cells. Rock inhibition decreases c-myc gene expression without affecting the E2F and c-Jun pathways. Loss-of-function studies demonstrate that c-Myc is important for the blockage of cell-contact inhibition rather than for promoting the proliferation of Rho-transformed cells. However, c-Myc overexpression does not bypass the inhibition of cell transformation induced by Rock blockage, indicating that c-Myc is essential, but not sufficient, for Rock-dependent transformation. These results reveal the complexity of the genetic program orchestrated by the Rho subfamily and pinpoint protein networks that mediate different aspects of the malignant phenotype of Rho-transformed cells.

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

Accession codes

Accessions

GenBank/EMBL/DDBJ

References

  • Barsyte-Lovejoy D, Mao DY, Penn LZ . (2004). c-Myc represses the proximal promoters of GADD45a and GADD153 by a post-RNA polymerase II recruitment mechanism. Oncogene 23: 3481–3486.

    Article  CAS  Google Scholar 

  • Bolstad BM, Collin F, Simpson KM, Irizarry RA, Speed TP . (2004). Experimental design and low-level analysis of microarray data. Int Rev Neurobiol 60: 25–58.

    Article  CAS  Google Scholar 

  • Budzyn K, Marley PD, Sobey CG . (2006). Targeting Rho and Rho-kinase in the treatment of cardiovascular disease. Trends Pharmacol Sci 27: 97–104.

    Article  CAS  Google Scholar 

  • Calvano SE, Xiao W, Richards DR, Felciano RM, Baker HV, Cho RJ et al. (2005). A network-based analysis of systemic inflammation in humans. Nature 437: 1032–1037.

    Article  CAS  Google Scholar 

  • Chen C, Nussenzweig A, Guo M, Kim D, Li GC, Ling CC . (1996). Down-regulation of gadd153 by c-myc in rat fibroblasts and its effect on cell growth and radiation-induced apoptosis. Oncogene 13: 1659–1665.

    CAS  PubMed  Google Scholar 

  • Etienne-Manneville S, Hall A . (2002). Rho GTPases in cell biology. Nature 420: 629–635.

    Article  CAS  Google Scholar 

  • Gautier L, Cope L, Bolstad BM, Irizarry RA . (2004). affy–analysis of affymetrix genechip data at the probe level. Bioinformatics 20: 307–315.

    Article  CAS  Google Scholar 

  • Gentleman RC, Carey VJ, Bates DM, Bolstad B, Dettling M, Dudoit S et al. (2004). Bioconductor: open software development for computational biology and bioinformatics. Genome Biol 5: R80.

    Article  Google Scholar 

  • Hakem A, Sanchez-Sweatman O, You-Ten A, Duncan G, Wakeham A, Khokha R et al. (2005). RhoC is dispensable for embryogenesis and tumor initiation but essential for metastasis. Genes Dev 19: 1974–1979.

    Article  CAS  Google Scholar 

  • Hill CS, Wynne J, Treisman R . (1995). The Rho family GTPases RhoA, Rac1, and CDC42Hs regulate transcriptional activation by SRF. Cell 81: 1159–1170.

    Article  CAS  Google Scholar 

  • Jaffe AB, Hall A . (2005). Rho GTPases: biochemistry and biology. Annu Rev Cell Dev Biol 21: 247–269.

    Article  CAS  Google Scholar 

  • Leng X, Connell-Crowley L, Goodrich D, Harper JW . (1997). S-Phase entry upon ectopic expression of G1 cyclin-dependent kinases in the absence of retinoblastoma protein phosphorylation. Curr Biol 7: 709–712.

    Article  CAS  Google Scholar 

  • Liu AX, Rane N, Liu JP, Prendergast GC . (2001). RhoB is dispensable for mouse development, but it modifies susceptibility to tumor formation as well as cell adhesion and growth factor signaling in transformed cells. Mol Cell Biol 21: 6906–6912.

    Article  CAS  Google Scholar 

  • Marinissen MJ, Chiariello M, Tanos T, Bernard O, Narumiya S, Gutkind JS . (2004). The small GTP-binding protein RhoA regulates c-jun by a ROCK-JNK signaling axis. Mol Cell 14: 29–41.

    Article  CAS  Google Scholar 

  • Montaner S, Perona R, Saniger L, Lacal JC . (1998). Multiple signalling pathways lead to the activation of the nuclear factor kappaB by the Rho family of GTPases. J Biol Chem 273: 12779–12785.

    Article  CAS  Google Scholar 

  • Montaner S, Perona R, Saniger L, Lacal JC . (1999). Activation of serum response factor by RhoA is mediated by the nuclear factor-kappaB and C/EBP transcription factors. J Biol Chem 274: 8506–8515.

    Article  CAS  Google Scholar 

  • Parrish RS, Spencer III HJ . (2004). Effect of normalization on significance testing for oligonucleotide microarrays. J Biopharm Stat 14: 575–589.

    Article  Google Scholar 

  • Perona R, Montaner S, Saniger L, Sanchez-Perez I, Bravo R, Lacal JC . (1997). Activation of the nuclear factor-kappaB by Rho, CDC42, and Rac-1 proteins. Genes Dev 11: 463–475.

    Article  CAS  Google Scholar 

  • Riento K, Ridley AJ . (2003). Rocks: multifunctional kinases in cell behaviour. Nat Rev Cell Biol 4: 446–456.

    Article  CAS  Google Scholar 

  • Sahai E, Ishizaki T, Narumiya S, Treisman R . (1999). Transformation mediated by RhoA requires activity of ROCK kinases. Curr Biol 9: 136–145.

    Article  CAS  Google Scholar 

  • Schuebel KE, Movilla N, Rosa JL, Bustelo XR . (1998). Phosphorylation-dependent and constitutive activation of Rho proteins by wild-type and oncogenic Vav-2. EMBO J 17: 6608–6621.

    Article  CAS  Google Scholar 

  • Simpson KJ, Dugan AS, Mercurio AM . (2004). Functional analysis of the contribution of RhoA and RhoC GTPases to invasive breast carcinoma. Cancer Res 64: 8694–8701.

    Article  CAS  Google Scholar 

  • Teramoto H, Malek RL, Behbahani B, Castellone MD, Lee NH, Gutkind JS . (2003). Identification of H-Ras, RhoA, Rac1 and Cdc42 responsive genes. Oncogene 22: 2689–2697.

    Article  CAS  Google Scholar 

  • Uehata M, Ishizaki T, Satoh H, Ono T, Kawahara T, Morishita T et al. (1997). Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension. Nature 389: 990–994.

    Article  CAS  Google Scholar 

  • van der Eb AJ, Graham FL . (1980). Assay of transforming activity of tumor virus DNA. Methods Enzymol 65: 826–839.

    Article  CAS  Google Scholar 

  • Watnick RS, Cheng YN, Rangarajan A, Ince TA, Weinberg RA . (2003). Ras modulates Myc activity to repress thrombospondin-1 expression and increase tumor angiogenesis. Cancer Cell 3: 219–231.

    Article  CAS  Google Scholar 

  • Wheeler AP, Ridley AJ . (2004). Why three Rho proteins? RhoA, RhoB, RhoC, and cell motility. Exp Cell Res 301: 43–49.

    Article  CAS  Google Scholar 

  • Wu M, Wu ZF, Kumar-Sinha C, Chinnaiyan A, Merajver SD . (2004). RhoC induces differential expression of genes involved in invasion and metastasis in MCF10A breast cells. Breast Cancer Res Treat 84: 3–12.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the Ramón Areces Foundation, the NCI/NIH (5RO1-CA73735-10) and the Spanish Ministry of Education and Science Special Action on Genomics and Proteomics (GEN2003-20239-C06-01). IMB was supported by a Spanish Ministry of Education and Science FPU fellowship (FP2000-6489). FN was partially supported by a Ernst Schering Foundation fellowship. The genomic data of this work are deposited in the NCBI Gene Expression Omnibus database (Accession number: GSE5913).

Author information

Author notes

  1. F Núñez

    Present address: 2Current address: Institut de Reserca, Hospital Vall d’Hebron, Psg. Vall d’Hebron 119-129, Barcelona, E-8035, Spain.,

Authors and Affiliations

  1. Centro de Investigación del Cáncer and Instituto de Biología Molecular y Celular del Cáncer (IBMCC), CSIC-University of Salamanca, Campus Unamuno, Salamanca, Spain

    I M Berenjeno, F Núñez & X R Bustelo

Authors
  1. I M Berenjeno
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. F Núñez
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. X R Bustelo
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to X R Bustelo.

Additional information

Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Berenjeno, I., Núñez, F. & Bustelo, X. Transcriptomal profiling of the cellular transformation induced by Rho subfamily GTPases. Oncogene 26, 4295–4305 (2007). https://doi.org/10.1038/sj.onc.1210194

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1210194

Keywords

This article is cited by

Search

Advanced search

Quick links