Use of a temperature-sensitive mutant to define the biological effects of the p210BCR-ABL tyrosine kinase on proliferation of a factor-dependent murine myeloid cell line

Nadia Carlesso, James D. Griffin, Brian Druker

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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, p210BCR-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 p210BCR-ABL function we constructed a model system in which the tyrosine kinase activity of p210BCR-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 p210BCR-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 p210BCR-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 1994

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Myeloid Cells
Protein-Tyrosine Kinases
Cell Line
Temperature
Interleukin-3
Tyrosine
Intercellular Signaling Peptides and Proteins
Phosphotransferases
Amino Acids
Chromosomes, Human, Pair 22
Philadelphia Chromosome
Chromosomes, Human, Pair 9
Proteins
Amino Acid Substitution
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Oncogenes
Exons
Cell Death
Phosphorylation
Mutation

ASJC Scopus subject areas

  • Cancer Research
  • Genetics
  • Molecular Biology

Cite this

Use of a temperature-sensitive mutant to define the biological effects of the p210BCR-ABL tyrosine kinase on proliferation of a factor-dependent murine myeloid cell line. / Carlesso, Nadia; Griffin, James D.; Druker, Brian.

In: Oncogene, Vol. 9, No. 1, 01.1994, p. 149-156.

Research output: Contribution to journalArticle

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