Role of p21 RAS in p210 bcr-abl transformation of murine myeloid cells

Blood. 1993 Sep 15;82(6):1838-47.

Abstract

The p21 RAS product has been implicated as part of the downstream signaling of certain nonreceptor tyrosine kinase oncogenes and several growth factor receptor-ligand interactions. We have reported that the chronic myelogenous leukemia oncogene p210 bcr-abl transforms a growth-factor-dependent myeloid cell line NFS/N1.H7 to interleukin-3 (IL-3) independence. In these p210 bcr-abl-transformed cells (H7 bcr-abl.A54) and in two other murine myeloid cell lines transformed to IL-3 independence by p210 bcr-abl, endogenous p21 RAS is activated as determined by an elevated ratio of associated guanosine triphosphate (GTP)/guanosine diphosphate (GDP), assayed by thin-layer chromatography of the nucleotides eluted from p21 RAS after immunoprecipitation with the Y13-259 antibody. Treatment of p210 bcr-abl-transformed cells with a specific tyrosine kinase inhibitor herbimycin A resulted in diminished tyrosine phosphorylation of p210 bcr-abl and associated proteins, without major reduction in expression of the p210 bcr-abl protein itself. Inhibition of p210 bcr-abl-dependent tyrosine phosphorylation resulted in a reduction of active p21RAS-GTP complexes in the transformed cells, in diminished expression of the nuclear early response genes c-jun and c-fos, and in lower cellular proliferation rate. To further implicate p21 RAS in these functional events downstream of p210 bcr-abl tyrosine phosphorylation, we targeted G-protein function directly by limiting the availability of GTP with the inosine monophosphate dehydrogenase inhibitor, tiazofurin (TR). In p210 bcr-abl-transformed cells treated for 4 hours with TR, in which the levels of GTP were reduced by 50%, but GDP, guanosine monophosphate, and adenosine triphosphate (ATP) were unaffected, p210 bcr-abl tyrosine phosphorylation was at control levels. However, expression of c-fos and c-jun nuclear proto-oncogenes were strongly inhibited and p21 RAS activity was downregulated. These findings show that p210 bcr-abl transduces proliferative signals, in part, through downstream activation of p21 RAS. Furthermore, p21 RAS activity is linked to pathways that regulate c-jun and c-fos expression.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Benzoquinones
  • Blotting, Northern
  • Bone Marrow
  • Cell Line
  • Cell Transformation, Neoplastic*
  • DNA Probes
  • Fusion Proteins, bcr-abl / genetics
  • Fusion Proteins, bcr-abl / isolation & purification
  • Fusion Proteins, bcr-abl / metabolism*
  • Gene Expression
  • Genes, ras
  • Guanosine Diphosphate / metabolism
  • Guanosine Triphosphate / metabolism
  • Kinetics
  • Lactams, Macrocyclic
  • Mice
  • Oncogenes
  • Phosphorylation
  • Protein-Tyrosine Kinases / isolation & purification
  • Protein-Tyrosine Kinases / metabolism*
  • Proto-Oncogene Proteins p21(ras) / genetics
  • Proto-Oncogene Proteins p21(ras) / isolation & purification
  • Proto-Oncogene Proteins p21(ras) / metabolism*
  • Quinones / pharmacology
  • RNA / genetics
  • RNA / isolation & purification
  • Rifabutin / analogs & derivatives
  • Signal Transduction
  • Transcription, Genetic

Substances

  • Benzoquinones
  • DNA Probes
  • Lactams, Macrocyclic
  • Quinones
  • Guanosine Diphosphate
  • Rifabutin
  • RNA
  • herbimycin
  • Guanosine Triphosphate
  • Protein-Tyrosine Kinases
  • Fusion Proteins, bcr-abl
  • Proto-Oncogene Proteins p21(ras)