title: [Detection of a non-standard mutation in the ret protoncogene by site directed mutagenesis].
abstract: MEN2A is an autosomic dominant disease, characterized by medullary thyroid cancer, pheochromocytoma and parathyroid hyperplasia. Mutations in the ret proto-oncogene are associated with this disease, with almost 100% of penetrance. The gene, situated on chromosome 10q11.2, codes for a transmembrane protein with a tyrosinkinase-like receptor function. Mutations that affect its extracellular domain, stimulate spontaneous homodimerization and elevate the basal tyrosinkinase activity. The codon 634 of the gene is considered a hot-spot site, since it is mutated in 85% of the MEN2A families. Our group developed in 2002 an indirect and costless strategy to detect alterations in this site. We present a family suspected of having MEN2A. We applied our PCR based indirect strategy on the DNA of the index patient and found that there was no mutation in that site. Posterior sequencing of exon 10 and 11 confirmed that the mutation affecting this family was in codon 611. Thus, we developed a new costless family-specific strategy based on mutagenic PCR and enzymatic cuts to diagnose all the family members. A seven-year old boy with this mutation was preventively thyroidectomized. In this way, combining the indirect methodology for codon 634 previously developed by our group, and a posterior family-specific mutation detection strategy, we were able to diagnose and intervene presymptomatically the family members, avoiding sending all the samples to foreign centers.
authors.affiliation: Laboratorio de Biología Celular y Molecular, IHEM-CONICET, Mendoza, Argentina.
authors.forename: Sebastián
authors.initials: S
authors.lastname: Real
authors.forename: Laura
authors.initials: L
authors.lastname: Gómez
authors.forename: Héctor
authors.initials: H
authors.lastname: Perinetti
authors.forename: Luis S
authors.initials: LS
authors.lastname: Mayorga
authors.forename: Eduardo
authors.initials: E
authors.lastname: Pusiol
authors.forename: María
authors.initials: M
authors.lastname: Roqué
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D008297:Male; D018813:Multiple Endocrine Neoplasia Type 2a / Q000175:diagnosis / Q000235:genetics*; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D009154:Mutation; D010375:Pedigree; D016133:Polymerase Chain Reaction; D000090063:Proto-Oncogene Mas; D051096:Proto-Oncogene Proteins c-ret / Q000235:genetics*
chemical_list: C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D051096:Proto-Oncogene Proteins c-ret; C099282:RET protein, human