Use of a temperature-sensitive mutant to define the biological effects of the p210(BCR-ABL) tyrosine kinase on proliferation of a factor-dependent murine myeloid cell line

N. Carlesso, J. D. Griffin, B. J. Druker

Research output: Contribution to journalArticle

76 Scopus citations

Abstract

The Philadelphia chromosome, detected in virtually all cases of chronic myelogenous leukemia, is formed by a reciprocal translocation between chromosomes 9 and 22 that fuses BCR encoded sequences upstream of exon 2 of c-ABL. This oncogene produces a fusion protein, p210(BCR-ABL) in which the ABL tyrosine kinase activity is elevated. This elevated kinase activity is essential for transformation, but the mechanisms involved are unknown To investigate p210(BCR-ABL) function we constructed a model system in which the tyrosine kinase activity of p210(BCR-ABL) was inducible. Two amino acid substitutions, Arg to His at amino acid 457 and Tyr to His at amino acid 469 of c-abl, modeled on mutations known to render v-src temperature-sensitive for tyrosine kinase activity, were introduced into p210(BCR-ABL). This mutant was characterized in an IL-3 growth factor dependent murine myeloid cell line, 32Dc13. Cell lines expressing the temperature-sensitive mutant remained factor dependent at the non-permissive temperature, but at the permissive temperature displayed a marked reduction in cell death in the absence of growth factor and an exaggerated proliferative response to low levels of IL-3. Both the kinase activity of the mutant and the levels of tyrosine phosphorylated proteins are increased in the temperature-sensitive mutant at the permissive temperature. Further, tyrosine phosphorylation of potential substrates of the p210(BCR-ABL) tyrosine kinase, p120 rasGAP and its associated proteins of p190 and p62, only occurs at the permissive temperature in cells expressing the temperature-sensitive mutant.

Original languageEnglish (US)
Pages (from-to)149-156
Number of pages8
JournalOncogene
Volume9
Issue number1
StatePublished - Jan 1 1994
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cancer Research

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