Abstract
We have introduced three Hirschsprung (HSCR) mutations localized in the tyrosine kinase domain of RET into the RET/PTC2 chimaeric oncogene which is capable of transforming NIH3T3 mouse fibroblasts and of differentiating pC12 rat pheochromocytoma cells. The three HSCR mutations abolished the biological activity of RET/PTC2 in both cell types and significantly decreased its tyrosine phosphorylation. By contrast, a rare polymorphism in exon 18 does not alter the transforming capability of RET/PTC2 or its tyrosine phosphorylation. These data suggest a loss of function effect of HSCR mutations which might act through a dominant negative mechanism. Our model system is therefore capable of discriminating between causative HSCR mutations and rare polymorphisms in the tyrosine kinase domain of RET.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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3T3 Cells
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Animals
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Base Sequence
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Cell Differentiation
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Cell Transformation, Neoplastic
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Cyclic AMP-Dependent Protein Kinases / genetics
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Cyclic AMP-Dependent Protein Kinases / metabolism*
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Drosophila Proteins*
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Exons
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Genetic Complementation Test
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HeLa Cells
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Hirschsprung Disease / genetics*
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Humans
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Mice
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Molecular Sequence Data
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Mutagenesis, Site-Directed
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Mutation*
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PC12 Cells
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Phosphorylation
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Precipitin Tests
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Proto-Oncogene Proteins / genetics
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Proto-Oncogene Proteins / physiology*
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Proto-Oncogene Proteins c-ret
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Rats
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Receptor Protein-Tyrosine Kinases / genetics
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Receptor Protein-Tyrosine Kinases / physiology*
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / physiology
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Transfection
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Tyrosine / metabolism
Substances
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Drosophila Proteins
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Proto-Oncogene Proteins
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Recombinant Fusion Proteins
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Tyrosine
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Proto-Oncogene Proteins c-ret
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Receptor Protein-Tyrosine Kinases
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Ret protein, Drosophila
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Ret protein, mouse
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Ret protein, rat
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Cyclic AMP-Dependent Protein Kinases