title: Detection of thyroid-stimulating hormone receptor and Gsalpha mutations: in 75 toxic thyroid nodules by denaturing gradient gel electrophoresis.
abstract: The actual frequency of constitutively activating thyrotropin receptor or Gsalpha mutations in toxic thyroid nodules (TTNs) remains controversial as considerable variation in the prevalence of these mutations has been reported. We studied a series of 75 consecutive TTNs and performed mutation screening by the more sensitive method of denaturing gradient gel electrophoresis (DGGE) in addition to direct sequencing. Furthermore, the likelihood of somatic mutations occurring in genes other than that for the thyroid-stimulating hormone receptor (TSHR) and exons 7-9 of the Gsalpha protein gene was determined by clonality analysis of TTNs, which did not harbor mutations in the investigated genes. In 43 of 75 TTNs (57%) constitutively active TSHR mutations were identified. Six TSHR mutations were detected only by DGGE, underlining the importance of a sensitive screening method. Novel, constitutively activating mutations were identified at positions 425 (Ser-->Leu) and 512 (Leu-->Glu/Arg). Furthermore, a new base substitution was detected at position Pro639Ala (CCA-->GCA). Ten of 20 TSHR or Gsalpha mutation negative cases (50%) showed nonrandom X-chromosome inactivation, indicating clonal origin. In conclusion, somatic, constitutively activating TSHR mutations appear to be a major cause of TTNs (57%), while mutations in Gsalpha play a minor role (3%). The mutation negative but clonal cases indicate a probable involvement of somatic mutations other than in the TSH receptor or Gsalpha genes as the molecular cause of these hot nodules.
authors: Department of Internal Medicine III, University of Leipzig, Germany.
authors: B
authors: B
authors: Trülzsch
authors: K
authors: K
authors: Krohn
authors: P
authors: P
authors: Wonerow
authors: S
authors: S
authors: Chey
authors: H P
authors: HP
authors: Holzapfel
authors: F
authors: F
authors: Ackermann
authors: D
authors: D
authors: Führer
authors: R
authors: R
authors: Paschke
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D001482:Base Composition; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods; D004303:Dosage Compensation, Genetic; D004591:Electrophoresis, Polyacrylamide Gel; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000235:genetics*; D005787:Gene Frequency; D006801:Humans; D009154:Mutation / Q000235:genetics*; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction; D011989:Receptors, Thyrotropin / Q000737:chemistry / Q000235:genetics*; D016606:Thyroid Nodule / Q000235:genetics*
chemical_list: D003062:Codon; D011989:Receptors, Thyrotropin; D019205:GTP-Binding Protein alpha Subunits, Gs