Quantitative determination of Rap 1 activation in cyclic nucleotide-treated HL-60 leukemic cells: lack of Rap 1 activation in variant cells
| Autor: | Renate B. Pilz, F. C. Von Lintig, Gerry R. Boss |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Cancer Research
Recombinant Fusion Proteins Drug Resistance Hamster HL-60 Cells Biology Kidney Second Messenger Systems Cell Line Cyclic nucleotide chemistry.chemical_compound Cricetinae Cyclic AMP Genetics Animals Guanine Nucleotide Exchange Factors Humans Nucleotide Protein kinase A Cyclic GMP Molecular Biology chemistry.chemical_classification Mesocricetus fungi rap1 GTP-Binding Proteins Cell Differentiation Thionucleotides Neoplasm Proteins Enzyme Activation body regions Enzyme chemistry Biochemistry Cell culture Guanosine Triphosphate sense organs Signal transduction Quantitative analysis (chemistry) |
| Zdroj: | Oncogene. 19:4029-4034 |
| ISSN: | 1476-5594 0950-9232 |
| DOI: | 10.1038/sj.onc.1203741 |
| Popis: | We have previously isolated variant HL-60 cells that are resistant to cGMP-induced differentiation and showed that they are deficient in proteolytic cleavage and/or carboxyl methylation of Rap 1A (J. Biol. Chem. 269, 32155 - 32161, 1994 and Oncogene 17, 2211 - 2233, 1998). We have now developed an enzyme-based method for assessing Rap 1 activation which is quantitative and provides a measurement of the per cent of Rap molecules in the active GTP-bound state. Using this method, we show that cAMP and cGMP analogs activate Rap 1 in parental HL-60 cells but not in the variant cells and that H-89, a cAMP-dependent protein kinase inhibitor, has no effect on cAMP-induced Rap 1 activation in parental cells. Thus, cAMP activation of Rap 1 in HL-60 cells is likely through a cAMP-regulated guanine nucleotide exchange factor (cAMP-GEF) and since cAMP does not activate Rap 1 in the variant cells, the data suggest that full post-translational processing of Rap 1 is necessary for cAMP-GEF activation of Rap 1. Activation of Rap 1 by cGMP analogs has not been previously found and suggests possible cross-talk between the NO/cGMP signal transduction pathway and Rap 1 signaling. Oncogene (2000) 19, 4029 - 4034. |
| Databáze: | OpenAIRE |
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