Expression of beta-arrestins in toxic and cold thyroid nodules.
beta-Arrestins mediate agonist dependent desensitization of G protein-coupled receptors. Somatic TSH receptor mutations were identified in the majority of hot thyroid nodules. When transiently overexpressed in COS 7 cells these mutations resulted in constitutive activation of the cAMP pathway. However, the in vivo mechanisms and the in vivo desensitization of these TSH receptor mutations are unknown. Moreover, constitutively activated beta-adrenergic receptors are known to be constitutively desensitized. Therefore, we investigated the expression of beta-arrestins in toxic thyroid nodules (TTNs) with and without somatic TSH receptor mutation and in cold thyroid nodules (CTNs) by Western blotting and ELISA. Expression of beta-arrestin 2 was increased in all TTNs while beta-arrestin 2 expression was decreased in CTNs compared to their corresponding surrounding tissue. The mean beta-arrestin 1 expression was unchanged in the cytosol of TTNs, in membranes and cytosol of CTNs and decreased in the membranes of TTNs compared to their surrounding tissue. Transient coexpression of beta-arrestins 1 or 2 with the TSH receptor in HEK 293 cells and subsequent determination of cAMP showed that in vitro both beta-arrestins interact with the TSH receptor and are able to desensitize the receptor. The increased beta-arrestin 2 expression in TTNs and the desensitization of the TSH receptor by beta-arrestin 2 in vitro suggest that the beta-arrestin 2 expression is cAMP dependent and that beta-arrestin 2 very likely desensitizes the constitutively activated TSH receptor in toxic thyroid nodules.
Voigt
C
C
III, Medical Department, University of Leipzig, Ph. Rosenthal Str. 27, D-04103, Leipzig, Germany.
Holzapfel
H
H
Paschke
R
R
D019390:Arrestins / Q000096:biosynthesis* / Q000235:genetics; D015153:Blotting, Western; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D009154:Mutation; D020033:Protein Isoforms; D011989:Receptors, Thyrotropin / Q000235:genetics / Q000378:metabolism; D016606:Thyroid Nodule / Q000378:metabolism* / Q000473:pathology; D013972:Thyrotropin / Q000378:metabolism / Q000494:pharmacology; D014162:Transfection; D000071559:beta-Arrestin 1; D000071560:beta-Arrestin 2; D000071557:beta-Arrestins
C000605253:ARRB1 protein, human; C000605254:ARRB2 protein, human; D019390:Arrestins; D020033:Protein Isoforms; D011989:Receptors, Thyrotropin; D000071559:beta-Arrestin 1; D000071560:beta-Arrestin 2; D000071557:beta-Arrestins; D013972:Thyrotropin; D000242:Cyclic AMP