CRKL links p210BCR/ABL with paxillin in chronic myelogenous leukemia cells

Ravi Salgia, Naoki Uemura, Keiko Okuda, Jian Liang Li, Evan Pisick, Martin Sattler, Ron De Jong, Brian Druker, Nora Heisterkamp, Lan Bo Chen, John Groffen, James D. Griffin

Research output: Contribution to journalArticle

121 Citations (Scopus)

Abstract

The Philadelphia chromosome translocation generates a chimeric oncogene, BCR/ABL, which causes chronic myelogenous leukemia (CML). In primary neutrophils from patients with CML, the major novel tyrosine-phosphorylated protein is CRKL, an SH2-SH3-SH3 linker protein which has an overall homology of 60% to CRK, the human homologue of the v-crk oncogene product. Anti-CRKL immunoprecipitates from CML cells, but not normal cells, were found to contain p210BCR/ABL and c-ABL. Several other phosphoproteins were also detected in anti-CRKL immunoprecipitates, one of which has been identified as paxillin, a 68-kDa focal adhesion protein which we have previously shown to be phosphorylated by p210BCR/ABL. Using GST-CRKL fusion proteins, the SH3 domains of CRKL were found to bind c-ABL and p210BCR/ABL, while the SH2 domain of CRKL bound to paxillin, suggesting that CRKL could physically link p210BCR/ABL to paxillin. Paxillin contains three tyrosines in Tyr-X-X-Pro (Y-X-X-P) motifs consistent with amino acid sequences predicted to be optimal for binding to the CRKL-SH2 domain (at positions Tyr-31, Tyr-118, and Tyr-181). Each of these tyrosine residues was mutated to a phenylalanine residue, and in vitro binding assays indicated that paxillin tyrosines at positions 31 and 118, but not 181, are likely to be involved in CRKL-SH2 binding. These results suggest that the p210BCR/ABL oncogene may be physically linked to the focal adhesion-associated protein paxillin in hematopoietic cells by CRKL. This interaction could contribute to the known adhesive defects of CML cells.

Original languageEnglish (US)
Pages (from-to)29145-29150
Number of pages6
JournalJournal of Biological Chemistry
Volume270
Issue number49
StatePublished - Dec 8 1995

Fingerprint

Paxillin
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
src Homology Domains
Tyrosine
Focal Adhesions
Proteins
Oncogenes
Adhesion
Philadelphia Chromosome
Phosphoproteins
Oncogene Proteins
Chromosomes
Phenylalanine
Adhesives
Amino Acid Sequence
Assays
Neutrophils
Fusion reactions
Amino Acids
Defects

ASJC Scopus subject areas

  • Biochemistry

Cite this

Salgia, R., Uemura, N., Okuda, K., Li, J. L., Pisick, E., Sattler, M., ... Griffin, J. D. (1995). CRKL links p210BCR/ABL with paxillin in chronic myelogenous leukemia cells. Journal of Biological Chemistry, 270(49), 29145-29150.

CRKL links p210BCR/ABL with paxillin in chronic myelogenous leukemia cells. / Salgia, Ravi; Uemura, Naoki; Okuda, Keiko; Li, Jian Liang; Pisick, Evan; Sattler, Martin; De Jong, Ron; Druker, Brian; Heisterkamp, Nora; Chen, Lan Bo; Groffen, John; Griffin, James D.

In: Journal of Biological Chemistry, Vol. 270, No. 49, 08.12.1995, p. 29145-29150.

Research output: Contribution to journalArticle

Salgia, R, Uemura, N, Okuda, K, Li, JL, Pisick, E, Sattler, M, De Jong, R, Druker, B, Heisterkamp, N, Chen, LB, Groffen, J & Griffin, JD 1995, 'CRKL links p210BCR/ABL with paxillin in chronic myelogenous leukemia cells', Journal of Biological Chemistry, vol. 270, no. 49, pp. 29145-29150.
Salgia R, Uemura N, Okuda K, Li JL, Pisick E, Sattler M et al. CRKL links p210BCR/ABL with paxillin in chronic myelogenous leukemia cells. Journal of Biological Chemistry. 1995 Dec 8;270(49):29145-29150.
Salgia, Ravi ; Uemura, Naoki ; Okuda, Keiko ; Li, Jian Liang ; Pisick, Evan ; Sattler, Martin ; De Jong, Ron ; Druker, Brian ; Heisterkamp, Nora ; Chen, Lan Bo ; Groffen, John ; Griffin, James D. / CRKL links p210BCR/ABL with paxillin in chronic myelogenous leukemia cells. In: Journal of Biological Chemistry. 1995 ; Vol. 270, No. 49. pp. 29145-29150.
@article{57e9a164e3fe43f8ae93e40108e6f211,
title = "CRKL links p210BCR/ABL with paxillin in chronic myelogenous leukemia cells",
abstract = "The Philadelphia chromosome translocation generates a chimeric oncogene, BCR/ABL, which causes chronic myelogenous leukemia (CML). In primary neutrophils from patients with CML, the major novel tyrosine-phosphorylated protein is CRKL, an SH2-SH3-SH3 linker protein which has an overall homology of 60{\%} to CRK, the human homologue of the v-crk oncogene product. Anti-CRKL immunoprecipitates from CML cells, but not normal cells, were found to contain p210BCR/ABL and c-ABL. Several other phosphoproteins were also detected in anti-CRKL immunoprecipitates, one of which has been identified as paxillin, a 68-kDa focal adhesion protein which we have previously shown to be phosphorylated by p210BCR/ABL. Using GST-CRKL fusion proteins, the SH3 domains of CRKL were found to bind c-ABL and p210BCR/ABL, while the SH2 domain of CRKL bound to paxillin, suggesting that CRKL could physically link p210BCR/ABL to paxillin. Paxillin contains three tyrosines in Tyr-X-X-Pro (Y-X-X-P) motifs consistent with amino acid sequences predicted to be optimal for binding to the CRKL-SH2 domain (at positions Tyr-31, Tyr-118, and Tyr-181). Each of these tyrosine residues was mutated to a phenylalanine residue, and in vitro binding assays indicated that paxillin tyrosines at positions 31 and 118, but not 181, are likely to be involved in CRKL-SH2 binding. These results suggest that the p210BCR/ABL oncogene may be physically linked to the focal adhesion-associated protein paxillin in hematopoietic cells by CRKL. This interaction could contribute to the known adhesive defects of CML cells.",
author = "Ravi Salgia and Naoki Uemura and Keiko Okuda and Li, {Jian Liang} and Evan Pisick and Martin Sattler and {De Jong}, Ron and Brian Druker and Nora Heisterkamp and Chen, {Lan Bo} and John Groffen and Griffin, {James D.}",
year = "1995",
month = "12",
day = "8",
language = "English (US)",
volume = "270",
pages = "29145--29150",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "49",

}

TY - JOUR

T1 - CRKL links p210BCR/ABL with paxillin in chronic myelogenous leukemia cells

AU - Salgia, Ravi

AU - Uemura, Naoki

AU - Okuda, Keiko

AU - Li, Jian Liang

AU - Pisick, Evan

AU - Sattler, Martin

AU - De Jong, Ron

AU - Druker, Brian

AU - Heisterkamp, Nora

AU - Chen, Lan Bo

AU - Groffen, John

AU - Griffin, James D.

PY - 1995/12/8

Y1 - 1995/12/8

N2 - The Philadelphia chromosome translocation generates a chimeric oncogene, BCR/ABL, which causes chronic myelogenous leukemia (CML). In primary neutrophils from patients with CML, the major novel tyrosine-phosphorylated protein is CRKL, an SH2-SH3-SH3 linker protein which has an overall homology of 60% to CRK, the human homologue of the v-crk oncogene product. Anti-CRKL immunoprecipitates from CML cells, but not normal cells, were found to contain p210BCR/ABL and c-ABL. Several other phosphoproteins were also detected in anti-CRKL immunoprecipitates, one of which has been identified as paxillin, a 68-kDa focal adhesion protein which we have previously shown to be phosphorylated by p210BCR/ABL. Using GST-CRKL fusion proteins, the SH3 domains of CRKL were found to bind c-ABL and p210BCR/ABL, while the SH2 domain of CRKL bound to paxillin, suggesting that CRKL could physically link p210BCR/ABL to paxillin. Paxillin contains three tyrosines in Tyr-X-X-Pro (Y-X-X-P) motifs consistent with amino acid sequences predicted to be optimal for binding to the CRKL-SH2 domain (at positions Tyr-31, Tyr-118, and Tyr-181). Each of these tyrosine residues was mutated to a phenylalanine residue, and in vitro binding assays indicated that paxillin tyrosines at positions 31 and 118, but not 181, are likely to be involved in CRKL-SH2 binding. These results suggest that the p210BCR/ABL oncogene may be physically linked to the focal adhesion-associated protein paxillin in hematopoietic cells by CRKL. This interaction could contribute to the known adhesive defects of CML cells.

AB - The Philadelphia chromosome translocation generates a chimeric oncogene, BCR/ABL, which causes chronic myelogenous leukemia (CML). In primary neutrophils from patients with CML, the major novel tyrosine-phosphorylated protein is CRKL, an SH2-SH3-SH3 linker protein which has an overall homology of 60% to CRK, the human homologue of the v-crk oncogene product. Anti-CRKL immunoprecipitates from CML cells, but not normal cells, were found to contain p210BCR/ABL and c-ABL. Several other phosphoproteins were also detected in anti-CRKL immunoprecipitates, one of which has been identified as paxillin, a 68-kDa focal adhesion protein which we have previously shown to be phosphorylated by p210BCR/ABL. Using GST-CRKL fusion proteins, the SH3 domains of CRKL were found to bind c-ABL and p210BCR/ABL, while the SH2 domain of CRKL bound to paxillin, suggesting that CRKL could physically link p210BCR/ABL to paxillin. Paxillin contains three tyrosines in Tyr-X-X-Pro (Y-X-X-P) motifs consistent with amino acid sequences predicted to be optimal for binding to the CRKL-SH2 domain (at positions Tyr-31, Tyr-118, and Tyr-181). Each of these tyrosine residues was mutated to a phenylalanine residue, and in vitro binding assays indicated that paxillin tyrosines at positions 31 and 118, but not 181, are likely to be involved in CRKL-SH2 binding. These results suggest that the p210BCR/ABL oncogene may be physically linked to the focal adhesion-associated protein paxillin in hematopoietic cells by CRKL. This interaction could contribute to the known adhesive defects of CML cells.

UR - http://www.scopus.com/inward/record.url?scp=0028971466&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028971466&partnerID=8YFLogxK

M3 - Article

C2 - 7493940

AN - SCOPUS:0028971466

VL - 270

SP - 29145

EP - 29150

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 49

ER -