Inactivation of wild-type BCR/ABL tyrosine kinase in hematopoietic cells by mild hyperthermia

• Published in: Leukemia Vol. 14; no. 5; pp. 845 - 852
• Main Authors: JAIN, S. K, DE AOS, I, INAI, Y, LIU, F, VARTICOVSKI, L
• Format: Conference Proceeding Journal Article
• Language: English
• Published: London Nature Publishing 01.05.2000; Nature Publishing Group
• Subjects: 1-Phosphatidylinositol 3-kinase; 3T3 Cells; Abl gene; Abl protein; Amino acids; Animals; Antigens; BCR gene; BCR protein; Binding sites; Biological and medical sciences; Cell Line; Cell Line, Transformed; Cells; Fever; Fusion Proteins, bcr-abl - antagonists & inhibitors; Fusion Proteins, bcr-abl - genetics; Gene deletion; Genes, abl; Grb2 protein; Growth factors; Hematopoietic Stem Cells - cytology; Hematopoietic Stem Cells - enzymology; Hemopathies; Hot Temperature; Humans; Hyperthermia; In vivo methods and tests; Inactivation; Kinases; Kinetics; Leukemia; Medical sciences; Mice; Mutation; Oncogene Proteins v-abl - antagonists & inhibitors; Phosphatidylinositol 3-Kinases - metabolism; Phosphorylation; Phosphotyrosine; Protein-tyrosine kinase; Protein-Tyrosine Kinases - antagonists & inhibitors; Proteins; Radiotherapy. Instrumental treatment. Physiotherapy. Reeducation. Rehabilitation, orthophony, crenotherapy. Diet therapy and various other treatments (general aspects); Recombinant Proteins - antagonists & inhibitors; Recombinant Proteins - biosynthesis; Signal transduction; src Homology Domains; Temperature effects; Temperature-sensitive mutant; Transfection; Transformed cells; Tyrosine; United States > US; Human; Tyrosine; Phosphorylation; Enzyme; Transferases; Malignant hemopathy; Inactivation; Wild type; In vitro; Myeloproliferative syndrome; Chronic; Treatment; Aminoacid; C-Onc gene; Chronic myelocytic leukemia; Genetics; Protein-tyrosine kinase; Hybrid gene; Induced hyperthermia; Protooncogene
• ISSN: 0887-6924; 1476-5551
• DOI: 10.1038/sj.leu.2401759

Temperature-sensitive mutants of BCR/ABL tyrosine kinase have been extensively used to study the mechanisms of cell transformation and signal transduction. However, little is known about the effect of temperature on the activity of wild-type BCR/ABL gene product. In this study, we demonstrate that in vivo tyrosine kinase activity of p210, p190 BCR/ABL and v-abl are temperature-sensitive when expressed in hematopoietic cells and decline when temperature is raised 2 degrees C above normal range. In vitro tyrosine kinase activities of purified recombinant Abl and immunoprecipitated p210 BCR/ABL were also sensitive to increased temperature. Tyrosine phosphorylation of cellular proteins was markedly reduced in BCR/ABL transformed cells after 16 h at 39 degrees C, whereas the expression of BCR/ABL was unchanged. Temperature-induced downregulation of BCR/ABL kinase activity was reversible when cells were shifted back to 37 degrees C. The downregulation of Abl tyrosine kinase activity was not influenced by mutation or deletion of SH2 or SH3 domains or mutation of the GRB2 binding site. No increase in functional activity or expression of protein-tyrosine phosphatases, PTP-1B, SH-PTP1 or SH-PTP2 was detected in cells grown at 39 degrees C. Temperature-induced downregulation in tyrosine kinase activity correlated with decline in phosphotyrosine-associated PI 3-kinase whereas there was no change in growth factor independence of transformed hematopoietic cells. In conclusion, Abl tyrosine kinase has intrinsic sensitivity to temperature and BCR/ABL expressed in hematopoietic cells is downregulated by increasing temperature 2 degrees C. These observations provide a unique opportunity to identify cellular factor(s) which regulate BCR/ABL kinase in vivo and suggests possible novel treatment of CML by a mild hyperthermia.