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Transcriptional networks of knockout cell lines identify functional specificities of H-Ras and N-Ras: significant involvement of N-Ras in biotic and defense responses

Oncogene volume 26, pages 917–933 (2007)Cite this article

Abstract

We characterized differential gene expression profiles of fibroblast cell lines harboring single or double-homozygous null mutations in H-ras and N-ras. Whereas the expression level of the individual H-, N- and K-ras genes appeared unaffected by the presence or absence of the other ras loci, significant differences were observed between the expression profiles of cells missing N-ras and/or H-ras. Absence of N-ras produced much stronger effects than absence of H-ras over the profile of the cellular transcriptome. N-ras−/− and H-ras−/− fibroblasts displayed rather antagonistic expression profiles and the transcriptome of H-ras−/− cells was significantly closer to that of wild-type fibroblasts than to that of N-ras−/− cells. Classifying all differentially expressed genes into functional categories suggested specific roles for H-Ras and N-Ras. It was particularly striking in N-ras−/− cells the upregulation of a remarkable number of immunity-related genes, as well as of several loci involved in apoptosis. Reverse-phase protein array assays demonstrated in the same N-ras−/− cells the overexpression and nuclear migration of tyrosine phosphorylated signal transducer and activator of transcription 1 (Stat1) which was concomitant with transcriptional activation mediated by interferon-stimulated response elements. Significantly enhanced numbers of apoptotic cells were also detected in cultures of N-ras−/− cells. Our data support the notion that different Ras isoforms play functionally distinct cellular roles and indicate that N-Ras is significantly involved in immune modulation/host defense and apoptotic responses

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Acknowledgements

We thank R Pine (The Public Health Research Institute, Newark, NJ) for ISRE and GAS reporter constructs and E Petricoin (FDA/NIH Proteomic Facility, Bethesda, MD, USA) for support with protein array layout generation and analysis. This work was supported by Grants SAF2003-04177 and GEN2003-20239-C06-02 from MEC and Grant PI021570 from MSC, as well as institutional support from Red Temática C03/10 de Investigación de Centros de Cáncer (RTICCC) from ISCIII, MSC, Spain. CG was supported by Ramón y Cajal Program. EC was a predoctoral fellow from MEC.

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

    E Castellano, J De Las Rivas, C Guerrero & E Santos

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  1. E Castellano
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  2. J De Las Rivas
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  3. C Guerrero
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Castellano, E., De Las Rivas, J., Guerrero, C. et al. Transcriptional networks of knockout cell lines identify functional specificities of H-Ras and N-Ras: significant involvement of N-Ras in biotic and defense responses. Oncogene 26, 917–933 (2007). https://doi.org/10.1038/sj.onc.1209845

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  • DOI: https://doi.org/10.1038/sj.onc.1209845

Keywords

  • H-ras
  • N-ras
  • microarray
  • genomics
  • transcriptome
  • cluster analysis
  • differential gene expression

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