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A retroviral library genetic screen identifies IRF-2 as an inhibitor of N-ras-induced growth suppression in leukemic cells

Oncogene volume 24, pages 7327–7336 (2005)Cite this article

Abstract

Activating mutations of the N-ras gene occur at relatively high frequency in acute myeloid leukemia and myelodysplastic syndrome. Somewhat paradoxically, ectopic expression of activated N-ras in primary hematopoietic cells and myeloid cell lines (in some cases) can lead to inhibition of proliferation. Expression of mutant N-ras in murine hematopoietic stem/progenitor cells is sufficient to induce myeloid malignancies, but these pathologies occur with long latency. This suggests that mutations that disable the growth suppressive properties of N-ras in hematopoietic cells are required for the development of frank malignancy. In the present work, the growth suppression induced by a mutant N-ras gene in U937 myeloid cells was used as the basis to screen a retroviral cDNA library for genes that prevent mutant N-ras-induced growth suppression (i.e., putative cooperating oncogenes). This screen identified the gene for the transcription factor interferon regulatory factor-2 (IRF-2), and as confirmation of the screen, overexpression of this gene in U937 cells was shown to inhibit mutant N-ras-induced growth suppression. Also recovered from the screen were two truncated clones of an uncharacterized gene (interim official symbol: PP2135). Overexpression of this truncated PP2135 gene in U937 cells did not appear to abrogate mutant N-ras-induced growth suppression, but rather appeared to confer an increased sensitivity of U937 cells to retroviral infection, accounting for the recovery of this gene from the genetic screen.

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Accession codes

Accessions

GenBank/EMBL/DDBJ

Abbreviations

AML:

acute myeloid leukemia

EGFP:

enhanced green fluorescent protein

FACS:

fluorescence-activated cell sorting

GFPtpz:

yellow-shifted green fluorescent protein ‘topaz’ variant

HPC:

hematopoietic progenitor cell

HSC:

hematopoietic stem cell

IRF-1:

interferon regulatory factor-1

IRF-2:

interferon regulatory factor-2

IRF-E:

interferon regulatory factor element

ISRE:

interferon-stimulated responsive element

NOD/SCID:

nonobese diabetic/severe combined immunodeficient

VCM:

virus containing media

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Acknowledgements

We acknowledge facility support by, and helpful discussions at Johnson & Johnson Research. This work was supported by an NHMRC project grant, and GS was an NHMRC Principal Research Fellow during this work. TP was supported by an Australian Post-graduate Award.

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Authors and Affiliations

  1. School of Medical Sciences, University of New South Wales, Kensington, Sydney, Australia

    Toby Passioura & Geoff Symonds

  2. Children's Cancer Institute Australia, c/- Johnson and Johnson Research, Locked Bag 4555, Strawberry Hills, Sydney, 2012, NSW, Australia

    Sylvie Shen, Geoff Symonds & Alla Dolnikov

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  1. Toby Passioura
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Correspondence to Geoff Symonds.

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Passioura, T., Shen, S., Symonds, G. et al. A retroviral library genetic screen identifies IRF-2 as an inhibitor of N-ras-induced growth suppression in leukemic cells. Oncogene 24, 7327–7336 (2005). https://doi.org/10.1038/sj.onc.1208877

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