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 Druker

Research output: Contribution to journalArticle

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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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Proline
Tyrosine
Assays
Two-Hybrid System Techniques
Yeast
Cells
src Homology Domains
Sequence Deletion
Myeloid Cells
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Intercellular Signaling Peptides and Proteins
Neutrophils
Gels
Cell Line
Proteins

ASJC Scopus subject areas

  • Hematology

Cite this

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

Direct binding of CRKL to BCR-ABL is not required for BCR-ABL transformation. / Heaney, Conor; Kolibaba, Kathryn; Bhat, Arun; Oda, Tsukasa; Ohno, Sayuri; Fanning, Shane; Druker, Brian.

In: Blood, Vol. 89, No. 1, 01.01.1997, p. 297-306.

Research output: Contribution to journalArticle

Heaney, C, Kolibaba, K, Bhat, A, Oda, T, Ohno, S, Fanning, S & Druker, B 1997, 'Direct binding of CRKL to BCR-ABL is not required for BCR-ABL transformation', Blood, vol. 89, no. 1, pp. 297-306.
Heaney C, Kolibaba K, Bhat A, Oda T, Ohno S, Fanning S et al. Direct binding of CRKL to BCR-ABL is not required for BCR-ABL transformation. Blood. 1997 Jan 1;89(1):297-306.
Heaney, Conor ; Kolibaba, Kathryn ; Bhat, Arun ; Oda, Tsukasa ; Ohno, Sayuri ; Fanning, Shane ; Druker, Brian. / Direct binding of CRKL to BCR-ABL is not required for BCR-ABL transformation. In: Blood. 1997 ; Vol. 89, No. 1. pp. 297-306.
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