v-raf confers CSF-1 independent growth to a macrophage cell line and leads to immediate early gene expression without MAP-Kinase activation

Dirk Büscher, Persio Dello Sbarba, Robert A. Hipskind, Ulf R. Rapp, E. Richard Stanley, Manuela Baccarini

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Abstract

The BAC-1.2F5 macrophage cell line depends on CSF-1 for proliferation and survival. Phosphorylation and activation of the RAF-1 kinase are among the early events in CSF-1 signal transduction. To characterize the role of RAF-1 in CSF-1-induced proliferation, we overexpressed oncogenically activated RAF-1, cellular RAF-1 and RAF-1 kinase-defective mutant proteins in BAC-1.2F5 cells. We were unable to establish stable cell lines expressing either kinase-negative or full length RAF-1 proteins, implying that expression of these molecules is not tolerated in BAC-1.2F5 cells. Oncogenically activated RAF-1 induces CSF-1-independent growth in the absence of autocrine growth factor production. Autonomous growth is not associated with dedifferentiation, since v-raf-expressing macrophages perform the same immunological functions as control cells. Intriguingly, autonomous growth correlates with the suppression of CSF-1-mediated MAP-Kinase activation and with the low constitutive expression of a number of CSF-1-inducible genes, including fos, jun, ets2, and myc, but also the genes for the inflammatory cytokines TNFα and IL-1β. Many of these genes have AP-1 binding sites in their promoters, and the v-raf-expressing cells contain constitutive AP-1 binding activity. These data indicate that RAF-1, but not MAP-Kinase, is a key component in CSF-1 mitogenic signal transduction, and are consistent with a working hypothesis in which RAF-1 mediates transcriptional activation of genes via AP-1.

Original languageEnglish (US)
Pages (from-to)3323-3332
Number of pages10
JournalOncogene
Volume8
Issue number12
StatePublished - Dec 1993

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Immediate-Early Genes
Macrophage Colony-Stimulating Factor
Phosphotransferases
Macrophages
Gene Expression
Cell Line
Growth
Transcription Factor AP-1
Signal Transduction
fos Genes
myc Genes
Mutant Proteins
Interleukin-1
Transcriptional Activation
Genes
Intercellular Signaling Peptides and Proteins
Binding Sites
Phosphorylation
Cytokines

ASJC Scopus subject areas

  • Cancer Research
  • Genetics
  • Molecular Biology

Cite this

Büscher, D., Dello Sbarba, P., Hipskind, R. A., Rapp, U. R., Stanley, E. R., & Baccarini, M. (1993). v-raf confers CSF-1 independent growth to a macrophage cell line and leads to immediate early gene expression without MAP-Kinase activation. Oncogene, 8(12), 3323-3332.

v-raf confers CSF-1 independent growth to a macrophage cell line and leads to immediate early gene expression without MAP-Kinase activation. / Büscher, Dirk; Dello Sbarba, Persio; Hipskind, Robert A.; Rapp, Ulf R.; Stanley, E. Richard; Baccarini, Manuela.

In: Oncogene, Vol. 8, No. 12, 12.1993, p. 3323-3332.

Research output: Contribution to journalArticle

Büscher, D, Dello Sbarba, P, Hipskind, RA, Rapp, UR, Stanley, ER & Baccarini, M 1993, 'v-raf confers CSF-1 independent growth to a macrophage cell line and leads to immediate early gene expression without MAP-Kinase activation', Oncogene, vol. 8, no. 12, pp. 3323-3332.
Büscher D, Dello Sbarba P, Hipskind RA, Rapp UR, Stanley ER, Baccarini M. v-raf confers CSF-1 independent growth to a macrophage cell line and leads to immediate early gene expression without MAP-Kinase activation. Oncogene. 1993 Dec;8(12):3323-3332.
Büscher, Dirk ; Dello Sbarba, Persio ; Hipskind, Robert A. ; Rapp, Ulf R. ; Stanley, E. Richard ; Baccarini, Manuela. / v-raf confers CSF-1 independent growth to a macrophage cell line and leads to immediate early gene expression without MAP-Kinase activation. In: Oncogene. 1993 ; Vol. 8, No. 12. pp. 3323-3332.
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