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PMID: 10209040 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Signal transducer and activator of transcription (STAT)5 activation by BCR/ABL is dependent on intact Src homology (SH)3 and SH2 domains of BCR/ABL and is required for leukemogenesis.

The Journal of experimental medicine ·Vol. 189 ·No. 8 ·1999-04-19 ·Pages 1229-42

Nieborowska-Skorska M, Wasik MA, Slupianek A, Salomoni P, Kitamura T, Calabretta B, Skorski T

Abstract

Signal transducer and activator of transcription (STAT)5 is constitutively activated in BCR/ ABL-expressing cells, but the mechanisms and functional consequences of such activation are unknown. We show here that BCR/ABL induces phosphorylation and activation of STAT5 by a mechanism that requires the BCR/ABL Src homology (SH)2 domain and the proline-rich binding site of the SH3 domain. Upon expression in 32Dcl3 growth factor-dependent myeloid precursor cells, STAT5 activation-deficient BCR/ABL SH3+SH2 domain mutants functioned as tyrosine kinase and activated Ras, but failed to protect from apoptosis induced by withdrawal of interleukin 3 and/or serum and did not induce leukemia in severe combined immunodeficiency mice. In complementation assays, expression of a dominant-active STAT5B mutant (STAT5B-DAM), but not wild-type STAT5B (STAT5B-WT), in 32Dcl3 cells transfected with STAT5 activation-deficient BCR/ABL SH3+SH2 mutants restored protection from apoptosis, stimulated growth factor-independent cell cycle progression, and rescued the leukemogenic potential in mice. Moreover, expression of a dominant-negative STAT5B mutant (STAT5B-DNM) in 32Dcl3 cells transfected with wild-type BCR/ABL inhibited apoptosis resistance, growth factor-independent proliferation, and the leukemogenic potential of these cells. In retrovirally infected mouse bone marrow cells, expression of STAT5B-DNM inhibited BCR/ABL-dependent transformation. Moreover, STAT5B-DAM, but not STAT5B-WT, markedly enhanced the ability of STAT5 activation-defective BCR/ABL SH3+SH2 mutants to induce growth factor-independent colony formation of primary mouse bone marrow progenitor cells. However, STAT5B-DAM did not rescue the growth factor-independent colony formation of kinase-deficient K1172R BCR/ABL or the triple mutant Y177F+R522L+ Y793F BCR/ABL, both of which also fail to activate STAT5. Together, these data demonstrate that STAT5 activation by BCR/ABL is dependent on signaling from more than one domain and document the important role of STAT5-regulated pathways in BCR/ABL leukemogenesis.

MeSH Terms
Animals Apoptosis Bone Marrow Cells/metabolism Cell Cycle/genetics DNA Replication/genetics DNA-Binding Proteins/genetics Genes, abl/genetics Genes, ras/genetics Leukemia/genetics Mice Mice, SCID Milk Proteins Mutation Phosphoproteins/analysis Phosphorylation STAT5 Transcription Factor Signal Transduction/genetics Stem Cells/metabolism Trans-Activators/genetics Transcriptional Activation/genetics src Homology Domains/genetics
Chemicals
DNA-Binding Proteins Milk Proteins Phosphoproteins STAT5 Transcription Factor Stat5b protein, mouse Trans-Activators
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Nieborowska-Skorska M
Department of Microbiology and Immunology, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Wasik M A
Slupianek A
Salomoni P
Kitamura T
Calabretta B
Skorski T
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Article Info
Journal
The Journal of experimental medicine
Abbr.
J Exp Med
ISSN
0022-1007
DOI
10.1084/jem.189.8.1229
Published
1999-04-19
Pages
1229-42
Language
English
Region
United States
NLM ID
2985109R
PMCID
PMC2193033
Subset
IM
Grants
NCI NIH HHS · CA46782-10 · United States
NCI NIH HHS · R29-CA70815 · United States
External Links
PubMed source DOI: 10.1084/jem.189.8.1229 PMC Full Text View journal details
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