Dominant Negative Effect of Mutated Thyroid Stimulating Hormone Receptor (P556L) Causes Hypothyroidism in C.RF-Tshrhytwild Mice 英文参考文献.docVIP

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Dominant Negative Effect of Mutated Thyroid Stimulating Hormone Receptor (P556L) Causes Hypothyroidism in C.RF-Tshrhytwild Mice 英文参考文献.doc

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Dominant Negative Effect of Mutated Thyroid Stimulating Hormone Receptor (P556L) Causes Hypothyroidism in C.RF-Tshrhytwild Mice 英文参考文献

DominantNegativeEffectofMutatedThyroid StimulatingHormoneReceptor(P556L)Causes HypothyroidisminC.RF-Tshrhyt/wild Mice ToyoshiEndo*,TetsuroKobayashi TheThirdDepartmentofInternalMedicine,InterdisciplinaryGraduateSchoolofMedicineandEngineering,UniversityofYamanashi,ChuoCity,Yamanashi,Japan Abstract C.RF-Tshrhyt/hyt micehaveamutatedthyroidstimulatinghormonereceptor(P556L-TSHR)andthesemicedevelopsevere hypothyroidism. We found that C.RF-Tshrhyt/wild heterozygous mice are also in a hypothyroid state. Thyroid glands from C.RF-Tshrhyt/wild micearesmallerthanthosefromwild-typemice,and 125Iuptakeactivitiesoftheformeraresignificantly lowerthanthoseinthelatter.WhenTSHR(TSHR(W))andP556L-TSHR(TSHR(M))cDNAswereclonedandco-transfectedinto HEK293cells,thecellsretained 125I-TSHbindingactivity,butcAMPresponsetoTSHwasdecreasedtoabout20%ofHEK 293 cells transfected with TSHR(W) cDNA. When TSHR(W) and TSHR(M) were tagged with eCFP or eYFP, we observed fluorescenceresonanceenergytransfer(FRET)inHEK293cellsexpressingTSHR(W)-eCFPandTSHR(W)-eYFPintheabsence ofTSH,butnotinthepresenceofTSH.Incontrast,weobtainedFRETinHEK293cellsexpressingTSHR(W)-eCFPandTSHR (M)-eYFP,regardlessofthepresenceorabsenceofTSH.TheseresultssuggestthatP556LTSHRhasadominantnegative effect on TSHR(W) by impairing polymer to monomer dissociation, which decreases TSH responsiveness and induces hypothyroidisminC.RF-Tshrhyt/wild mice. Citation: Endo T, Kobayashi T (2012) Dominant Negative Effect of Mutated Thyroid Stimulating Hormone Receptor (P556L) Causes Hypothyroidism in C.RF- Tshrhyt/wild Mice.PLoSONE7(8):e42358.doi:10.1371/journal.pone.0042358 Editor:MarianLudgate,CardiffUniversity,UnitedKingdom ReceivedApril11,2012;AcceptedJuly4,2012;PublishedAugust16,2012 Copyright: ?2012Endo,Kobayashi.Thisisanopen-accessarticledistributedunderthetermsoftheCreativeCommonsAttributionLicense,whichpermits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:Theauthorshav