title: The Glu632-Leu633 deletion in cysteine rich domain of Ret induces constitutive dimerization and alters the processing of the receptor protein.
abstract: Mutations of the RET gene, encoding a receptor tyrosine kinase, have been associated with the inherited cancer syndromes MEN 2A and MEN 2B. They have also further been associated with both familial and sporadic medullary thyroid carcinomas. Missense mutations affecting cysteine residues within the extracellular domain of the receptor causes constitutive tyrosine kinase activation through the formation of disulfide-bonded homodimers. We have recently reported that a somatic 6 bp in-frame deletion, originally coding for Glu632-Leu633, potently activates the RET gene. This activation is increased with respect to the frequent MEN 2A-associated missense mutation Cys634Arg. This finding specifically correlated to the clinic behavior of the corresponding tumor, which was characterized by an unusually aggressive progression with both multiple and recurrent metastases. By examining the possibility that this deletion acts in a manner similar to cysteine substitution, we have analysed the molecular mechanism by which this oncogenic activation occurs. Phosphorylated dimers of the deleted Ret receptor were detected in immunoprecipitates separated under non-reducing conditions. Like other Cys point mutations, this 6 bp deletion affecting two amino acid residues between two adjacent Cys, is capable of activating the transforming ability of Ret by promoting receptor dimerization. These results suggest that alteration to cysteine residue position or pairing is capable of inducing ligand independent dimerization. Furthermore, we present data demonstrating that the processing and sorting of the Ret membrane receptor to the cell surface is affected by mutation type.
authors.affiliation: Division of Experimental Oncology A, Instituto Nazionale Tumori, 20133 Milan, Italy.
authors.forename: I
authors.initials: I
authors.lastname: Bongarzone
authors.forename: E
authors.initials: E
authors.lastname: Vigano
authors.forename: L
authors.initials: L
authors.lastname: Alberti
authors.forename: P
authors.initials: P
authors.lastname: Mondellini
authors.forename: M
authors.initials: M
authors.lastname: Uggeri
authors.forename: B
authors.initials: B
authors.lastname: Pasini
authors.forename: M G
authors.initials: MG
authors.lastname: Borrello
authors.forename: M A
authors.initials: MA
authors.lastname: Pierotti
mesh_terms: D016475:3T3 Cells / Q000187:drug effects; D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002471:Cell Transformation, Neoplastic / Q000187:drug effects; D003545:Cysteine / Q000378:metabolism*; D019281:Dimerization; D029721:Drosophila Proteins; D004721:Endoplasmic Reticulum / Q000378:metabolism; D018698:Glutamic Acid; D007930:Leucine; D008623:Mercaptoethanol / Q000494:pharmacology; D051379:Mice; D010766:Phosphorylation; D011134:Polysaccharides / Q000378:metabolism; D011233:Precipitin Tests; D011498:Protein Precursors / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D051096:Proto-Oncogene Proteins c-ret; D020794:Receptor Protein-Tyrosine Kinases / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D017384:Sequence Deletion; D055765:Shc Signaling Adaptor Proteins; D015398:Signal Transduction; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; D029721:Drosophila Proteins; D011134:Polysaccharides; D011498:Protein Precursors; D011506:Proteins; D011518:Proto-Oncogene Proteins; D055765:Shc Signaling Adaptor Proteins; C528034:Shc1 protein, mouse; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D018698:Glutamic Acid; D008623:Mercaptoethanol; D051096:Proto-Oncogene Proteins c-ret; D020794:Receptor Protein-Tyrosine Kinases; C416649:Ret protein, Drosophila; C494999:Ret protein, mouse; D007930:Leucine; D003545:Cysteine