Direct binding of CRKL to BCR-ABL is not required for BCR-ABL transformation

Conor Heaney, Kathryn Kolibaba, Arun Bhat, Tsukasa Oda, Sayuri Ohno, Shane Fanning, Brian J. Druker

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69 Scopus citations

Abstract

CRKL has previously been shown to be a major tyrosine phosphorylated protein in neutrophils of patients with BCR-ABL+ chronic myelogenous leukemia and in cell lines expressing BCR-ABL. CRKL and BCR-ABL form a complex as demonstrated by coimmunoprecipitation and are capable of a direct interaction in a yeast two-hybrid assay. We have mapped the site of interaction of CRKL and BCR-ABL to the amino terminal SH3 domain of CRKL with a proline rich region in the C-terminus of ABL. The proline-rich region was mutated and the effect of this deletion on BCR-ABL transforming function was assayed. Our data show that this deletion does not impair the ability of BCR- ABL to render myeloid cells factor independent for growth. In cells expressing the proline deletion mutation of BCR-ABL, CRKL is still tyrosine phosphorylated and forms a complex with BCR-ABL as demonstrated by coimmunoprecipitation. Our data suggest that the interaction between CRKL and the proline deletion mutant of BCR-ABL is an indirect interaction as CRKL does not interact directly with the proline deletion mutant of BCR-ABL in a gel overlay assay or in a yeast two-hybrid assay. Thus, a direct interaction of CRKL and BCR-ABL is not required for CRKL to become tyrosine phosphorylated by BCR-ABL and suggests that CRKL function may still be required for BCR-ABL function through an indirect interaction.

Original languageEnglish (US)
Pages (from-to)297-306
Number of pages10
JournalBlood
Volume89
Issue number1
StatePublished - Jan 1 1997

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ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

Cite this

Heaney, C., Kolibaba, K., Bhat, A., Oda, T., Ohno, S., Fanning, S., & Druker, B. J. (1997). Direct binding of CRKL to BCR-ABL is not required for BCR-ABL transformation. Blood, 89(1), 297-306.