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

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.
Mesh Terms:
Adaptor Proteins, Signal Transducing, Binding Sites, Cell Division, Cell Line, Cell Transformation, Neoplastic, Fusion Proteins, bcr-abl, Hematopoietic Stem Cells, Humans, Mutagenesis, Site-Directed, Nuclear Proteins, Phosphorylation, Protein Binding, Protein Processing, Post-Translational, Recombinant Fusion Proteins, Sequence Deletion, src Homology Domains
Blood
Date: Jan. 01, 1997
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