title: Comparative proteomic analysis to dissect differences in signal transduction in activating TSH receptor mutations in the thyroid.
abstract: In the thyroid, cAMP controls both thyroid growth and function. Gain-of-function mutations in the thyroid-stimulating hormone receptor (TSHR) lead to constitutive cAMP formation and are a major cause of autonomous thyroid adenomas. The impact of activating TSHR mutations on the signal transduction network of the thyrocyte is not fully understood. To gain more insights into constitutive TSHR signaling, rat thyrocytes (FRTL-5 cells) with stable expression of three activating TSHR mutants (mutTSHR: A623I, L629F and Del613-621), which differ in their functional characteristics in vitro, were analyzed by a quantitative proteomic approach and compared to the wild-type TSHR (WT-TSHR). This study revealed (1) differences in the expression of Rab proteins suggesting an increased TSHR internalization in mutTSHR but not in the WT-TSHR; (2) differential stimulation of PI3K/Akt signaling in mutTSHR vs. WT-TSHR cells, (3) activation of Epac, impairing short-time Akt phosphorylation in both, mutTSHR and WT-TSHR cells. Based on the analysis of global changes in protein expression patterns, our findings underline the complexity of gain-of-function TSHR signaling in thyrocytes, which extends beyond pure cAMP and/or IP formation. Moreover, evidence for augmented endocytosis in the mutTSHR, adds to a new concept of TSHR signaling in thyroid autonomy. Further studies are required to clarify whether the observed differences in Rab, PI3K and Epac signaling may contribute to differences in the phenotypic presentation, i.e. stimulation of function and growth of thyroid autonomy in vivo.
authors.affiliation: Clinic for Endocrinology and Nephrology, University of Leipzig, Liebigstr. 20, D-04413 Leipzig, Germany.
authors.forename: Kerstin
authors.initials: K
authors.lastname: Krause
authors.forename: Alexandra
authors.initials: A
authors.lastname: Boisnard
authors.forename: Christian
authors.initials: C
authors.lastname: Ihling
authors.forename: Marian
authors.initials: M
authors.lastname: Ludgate
authors.forename: Markus
authors.initials: M
authors.lastname: Eszlinger
authors.forename: Knut
authors.initials: K
authors.lastname: Krohn
authors.forename: Andrea
authors.initials: A
authors.lastname: Sinz
authors.forename: Dagmar
authors.initials: D
authors.lastname: Fuhrer
mesh_terms: D000818:Animals; D000242:Cyclic AMP / Q000378:metabolism; D006801:Humans; D020935:MAP Kinase Signaling System; D009154:Mutation; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D040901:Proteomics; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D051381:Rats; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D011989:Receptors, Thyrotropin / Q000235:genetics* / Q000378:metabolism* / Q000502:physiology; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology; D013961:Thyroid Gland / Q000166:cytology / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D043562:Receptors, G-Protein-Coupled; D011989:Receptors, Thyrotropin; D000242:Cyclic AMP; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt