The Pathogenic TSH β-Subunit Variant C105Vfs114X Causes a Modified Signaling Profile at TSHR

(1) Background: Central congenital hypothyroidism (CCH) is a rare endocrine disorder that can be caused by mutations in the &#946;-subunit of thyrotropin (<i>TSHB</i>). The <i>TSHB</i> mutation C105Vfs114X leads to isolated thyroid-stimulating-hormone-(TSH)-deficiency and...

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Main Authors: Laura Kalveram, Gunnar Kleinau, Kamila Szymańska, Patrick Scheerer, Adolfo Rivero-Müller, Annette Grüters-Kieslich, Heike Biebermann
Format: Article
Language:English
Published: MDPI AG 2019-11-01
Series:International Journal of Molecular Sciences
Subjects:
Online Access:https://www.mdpi.com/1422-0067/20/22/5564
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spelling doaj-98e240326f184b89b34f78bde5d324062020-11-24T21:51:05ZengMDPI AGInternational Journal of Molecular Sciences1422-00672019-11-012022556410.3390/ijms20225564ijms20225564The Pathogenic TSH β-Subunit Variant C105Vfs114X Causes a Modified Signaling Profile at TSHRLaura Kalveram0Gunnar Kleinau1Kamila Szymańska2Patrick Scheerer3Adolfo Rivero-Müller4Annette Grüters-Kieslich5Heike Biebermann6Institute of Experimental Pediatric Endocrinology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 13353 Berlin, GermanyInstitute of Medical Physics and Biophysics, Group Protein X-ray Crystallography and Signal Transduction, Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 10117 Berlin, GermanyDepartment of Biochemistry and Molecular Biology, Medical University of Lublin, 20-093 Lublin, PolandInstitute of Medical Physics and Biophysics, Group Protein X-ray Crystallography and Signal Transduction, Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 10117 Berlin, GermanyDepartment of Biochemistry and Molecular Biology, Medical University of Lublin, 20-093 Lublin, PolandInstitute of Experimental Pediatric Endocrinology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 13353 Berlin, GermanyInstitute of Experimental Pediatric Endocrinology, Charité-Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 13353 Berlin, Germany(1) Background: Central congenital hypothyroidism (CCH) is a rare endocrine disorder that can be caused by mutations in the &#946;-subunit of thyrotropin (<i>TSHB</i>). The <i>TSHB</i> mutation C105Vfs114X leads to isolated thyroid-stimulating-hormone-(TSH)-deficiency and results in a severe phenotype. The aim of this study was to gain more insight into the underlying molecular mechanism and the functional effects of this mutation based on two assumptions: a) the three-dimensional (3D) structure of TSH should be modified with the C105V substitution, and/or b) whether the C-terminal modifications lead to signaling differences. (2) Methods: wild-type (WT) and different mutants of hTSH were generated in human embryonic kidney 293 cells (HEK293 cells) and TSH preparations were used to stimulate thyrotropin receptor (TSHR) stably transfected into follicular thyroid cancer cells (FTC133-TSHR cells) and transiently transfected into HEK293 cells. Functional characterization was performed by determination of Gs, mitogen activated protein kinase (MAPK) and Gq/11 activation. (3) Results: The patient mutation C105Vfs114X and further designed TSH mutants diminished cyclic adenosine monophosphate (cAMP) signaling activity. Surprisingly, MAPK signaling for all mutants was comparable to WT, while none of the mutants induced PLC activation. (4) Conclusion: We characterized the patient mutation C105Vfs114X concerning different signaling pathways. We identified a strong decrease of cAMP signaling induction and speculate that this could, in combination with diverse signaling regarding the other pathways, accounting for the patient&#8217;s severe phenotype.https://www.mdpi.com/1422-0067/20/22/5564central congenital hypothyroidismg-protein coupled receptorsthyroid-stimulating hormonetshr
collection DOAJ
language English
format Article
sources DOAJ
author Laura Kalveram
Gunnar Kleinau
Kamila Szymańska
Patrick Scheerer
Adolfo Rivero-Müller
Annette Grüters-Kieslich
Heike Biebermann
spellingShingle Laura Kalveram
Gunnar Kleinau
Kamila Szymańska
Patrick Scheerer
Adolfo Rivero-Müller
Annette Grüters-Kieslich
Heike Biebermann
The Pathogenic TSH β-Subunit Variant C105Vfs114X Causes a Modified Signaling Profile at TSHR
International Journal of Molecular Sciences
central congenital hypothyroidism
g-protein coupled receptors
thyroid-stimulating hormone
tshr
author_facet Laura Kalveram
Gunnar Kleinau
Kamila Szymańska
Patrick Scheerer
Adolfo Rivero-Müller
Annette Grüters-Kieslich
Heike Biebermann
author_sort Laura Kalveram
title The Pathogenic TSH β-Subunit Variant C105Vfs114X Causes a Modified Signaling Profile at TSHR
title_short The Pathogenic TSH β-Subunit Variant C105Vfs114X Causes a Modified Signaling Profile at TSHR
title_full The Pathogenic TSH β-Subunit Variant C105Vfs114X Causes a Modified Signaling Profile at TSHR
title_fullStr The Pathogenic TSH β-Subunit Variant C105Vfs114X Causes a Modified Signaling Profile at TSHR
title_full_unstemmed The Pathogenic TSH β-Subunit Variant C105Vfs114X Causes a Modified Signaling Profile at TSHR
title_sort pathogenic tsh β-subunit variant c105vfs114x causes a modified signaling profile at tshr
publisher MDPI AG
series International Journal of Molecular Sciences
issn 1422-0067
publishDate 2019-11-01
description (1) Background: Central congenital hypothyroidism (CCH) is a rare endocrine disorder that can be caused by mutations in the &#946;-subunit of thyrotropin (<i>TSHB</i>). The <i>TSHB</i> mutation C105Vfs114X leads to isolated thyroid-stimulating-hormone-(TSH)-deficiency and results in a severe phenotype. The aim of this study was to gain more insight into the underlying molecular mechanism and the functional effects of this mutation based on two assumptions: a) the three-dimensional (3D) structure of TSH should be modified with the C105V substitution, and/or b) whether the C-terminal modifications lead to signaling differences. (2) Methods: wild-type (WT) and different mutants of hTSH were generated in human embryonic kidney 293 cells (HEK293 cells) and TSH preparations were used to stimulate thyrotropin receptor (TSHR) stably transfected into follicular thyroid cancer cells (FTC133-TSHR cells) and transiently transfected into HEK293 cells. Functional characterization was performed by determination of Gs, mitogen activated protein kinase (MAPK) and Gq/11 activation. (3) Results: The patient mutation C105Vfs114X and further designed TSH mutants diminished cyclic adenosine monophosphate (cAMP) signaling activity. Surprisingly, MAPK signaling for all mutants was comparable to WT, while none of the mutants induced PLC activation. (4) Conclusion: We characterized the patient mutation C105Vfs114X concerning different signaling pathways. We identified a strong decrease of cAMP signaling induction and speculate that this could, in combination with diverse signaling regarding the other pathways, accounting for the patient&#8217;s severe phenotype.
topic central congenital hypothyroidism
g-protein coupled receptors
thyroid-stimulating hormone
tshr
url https://www.mdpi.com/1422-0067/20/22/5564
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