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

Romeo A. Mandanas, David S. Leibowitz, Kamran Gharehbaghi, Tetsuzo Tauchi, Gem S. Burgess, Keisuke Miyazawa, Hiremagalur N. Jayaram, H. Boswell

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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 resuited in a reduction of active p21 RAS-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.

Original languageEnglish (US)
Pages (from-to)1838-1847
Number of pages10
JournalBlood
Volume82
Issue number6
StatePublished - Sep 15 1993

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Phosphorylation
Myeloid Cells
Guanosine Triphosphate
tiazofurin
Tyrosine
jun Genes
Guanosine
Diphosphates
Interleukin-3
Protein-Tyrosine Kinases
Oncogenes
Cells
Guanosine Monophosphate
Thin layer chromatography
Inosine Monophosphate
Growth Factor Receptors
Level control
Cell Line
GTP-Binding Proteins
Intercellular Signaling Peptides and Proteins

ASJC Scopus subject areas

  • Hematology

Cite this

Mandanas, R. A., Leibowitz, D. S., Gharehbaghi, K., Tauchi, T., Burgess, G. S., Miyazawa, K., ... Boswell, H. (1993). Role of p21 RAS in p210 bcr-abl transformation of murine myeloid cells. Blood, 82(6), 1838-1847.

Role of p21 RAS in p210 bcr-abl transformation of murine myeloid cells. / Mandanas, Romeo A.; Leibowitz, David S.; Gharehbaghi, Kamran; Tauchi, Tetsuzo; Burgess, Gem S.; Miyazawa, Keisuke; Jayaram, Hiremagalur N.; Boswell, H.

In: Blood, Vol. 82, No. 6, 15.09.1993, p. 1838-1847.

Research output: Contribution to journalArticle

Mandanas, RA, Leibowitz, DS, Gharehbaghi, K, Tauchi, T, Burgess, GS, Miyazawa, K, Jayaram, HN & Boswell, H 1993, 'Role of p21 RAS in p210 bcr-abl transformation of murine myeloid cells', Blood, vol. 82, no. 6, pp. 1838-1847.
Mandanas RA, Leibowitz DS, Gharehbaghi K, Tauchi T, Burgess GS, Miyazawa K et al. Role of p21 RAS in p210 bcr-abl transformation of murine myeloid cells. Blood. 1993 Sep 15;82(6):1838-1847.
Mandanas, Romeo A. ; Leibowitz, David S. ; Gharehbaghi, Kamran ; Tauchi, Tetsuzo ; Burgess, Gem S. ; Miyazawa, Keisuke ; Jayaram, Hiremagalur N. ; Boswell, H. / Role of p21 RAS in p210 bcr-abl transformation of murine myeloid cells. In: Blood. 1993 ; Vol. 82, No. 6. pp. 1838-1847.
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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 resuited in a reduction of active p21 RAS-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.",
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AU - Burgess, Gem S.

AU - Miyazawa, Keisuke

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AU - Boswell, H.

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N2 - 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 resuited in a reduction of active p21 RAS-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.

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