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PTH1R Mutants Found in Patients with Primary Failure of Tooth Eruption Disrupt G-Protein Signaling
AIM: Primary failure of tooth eruption (PFE) is causally linked to heterozygous mutations of the parathyroid hormone receptor (PTH1R) gene. The mutants described so far lead to exchange of amino acids or truncation of the protein that may result in structural changes of the expressed PTH1R. However,...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5127517/ https://www.ncbi.nlm.nih.gov/pubmed/27898723 http://dx.doi.org/10.1371/journal.pone.0167033 |
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author | Subramanian, Hariharan Döring, Frank Kollert, Sina Rukoyatkina, Natalia Sturm, Julia Gambaryan, Stepan Stellzig-Eisenhauer, Angelika Meyer-Marcotty, Philipp Eigenthaler, Martin Wischmeyer, Erhard |
author_facet | Subramanian, Hariharan Döring, Frank Kollert, Sina Rukoyatkina, Natalia Sturm, Julia Gambaryan, Stepan Stellzig-Eisenhauer, Angelika Meyer-Marcotty, Philipp Eigenthaler, Martin Wischmeyer, Erhard |
author_sort | Subramanian, Hariharan |
collection | PubMed |
description | AIM: Primary failure of tooth eruption (PFE) is causally linked to heterozygous mutations of the parathyroid hormone receptor (PTH1R) gene. The mutants described so far lead to exchange of amino acids or truncation of the protein that may result in structural changes of the expressed PTH1R. However, functional effects of these mutations have not been investigated yet. MATERIALS AND METHODS: In HEK293 cells, PTH1R wild type was co-transfected with selected PTH1R mutants identified in patients with PFE. The effects on activation of PTH-regulated intracellular signaling pathways were analyzed by ELISA and Western immunoblotting. Differential effects of wild type and mutated PTH1R on TRESK ion channel regulation were analyzed by electrophysiological recordings in Xenopus laevis oocytes. RESULTS: In HEK293 cells, activation of PTH1R wild type increases cAMP and in response activates cAMP-stimulated protein kinase as detected by phosphorylation of the vasodilator stimulated phosphoprotein (VASP). In contrast, the PTH1R mutants are functionally inactive and mutant PTH1R/Gly452Glu has a dominant negative effect on the signaling of PTH1R wild type. Confocal imaging revealed that wild type PTH1R is expressed on the cell surface, whereas PTH1R/Gly452Glu mutant is mostly retained inside the cell. Furthermore, in contrast to wild type PTH1R which substantially augmented K(+) currents of TRESK channels, coupling of mutated PTH1R to TRESK channels was completely abolished. CONCLUSIONS: PTH1R mutations affect intracellular PTH-regulated signaling in vitro. In patients with primary failure of tooth eruption defective signaling of PTH1R mutations is suggested to occur in dento-alveolar cells and thus may lead to impaired tooth movement. |
format | Online Article Text |
id | pubmed-5127517 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-51275172016-12-15 PTH1R Mutants Found in Patients with Primary Failure of Tooth Eruption Disrupt G-Protein Signaling Subramanian, Hariharan Döring, Frank Kollert, Sina Rukoyatkina, Natalia Sturm, Julia Gambaryan, Stepan Stellzig-Eisenhauer, Angelika Meyer-Marcotty, Philipp Eigenthaler, Martin Wischmeyer, Erhard PLoS One Research Article AIM: Primary failure of tooth eruption (PFE) is causally linked to heterozygous mutations of the parathyroid hormone receptor (PTH1R) gene. The mutants described so far lead to exchange of amino acids or truncation of the protein that may result in structural changes of the expressed PTH1R. However, functional effects of these mutations have not been investigated yet. MATERIALS AND METHODS: In HEK293 cells, PTH1R wild type was co-transfected with selected PTH1R mutants identified in patients with PFE. The effects on activation of PTH-regulated intracellular signaling pathways were analyzed by ELISA and Western immunoblotting. Differential effects of wild type and mutated PTH1R on TRESK ion channel regulation were analyzed by electrophysiological recordings in Xenopus laevis oocytes. RESULTS: In HEK293 cells, activation of PTH1R wild type increases cAMP and in response activates cAMP-stimulated protein kinase as detected by phosphorylation of the vasodilator stimulated phosphoprotein (VASP). In contrast, the PTH1R mutants are functionally inactive and mutant PTH1R/Gly452Glu has a dominant negative effect on the signaling of PTH1R wild type. Confocal imaging revealed that wild type PTH1R is expressed on the cell surface, whereas PTH1R/Gly452Glu mutant is mostly retained inside the cell. Furthermore, in contrast to wild type PTH1R which substantially augmented K(+) currents of TRESK channels, coupling of mutated PTH1R to TRESK channels was completely abolished. CONCLUSIONS: PTH1R mutations affect intracellular PTH-regulated signaling in vitro. In patients with primary failure of tooth eruption defective signaling of PTH1R mutations is suggested to occur in dento-alveolar cells and thus may lead to impaired tooth movement. Public Library of Science 2016-11-29 /pmc/articles/PMC5127517/ /pubmed/27898723 http://dx.doi.org/10.1371/journal.pone.0167033 Text en © 2016 Subramanian et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Subramanian, Hariharan Döring, Frank Kollert, Sina Rukoyatkina, Natalia Sturm, Julia Gambaryan, Stepan Stellzig-Eisenhauer, Angelika Meyer-Marcotty, Philipp Eigenthaler, Martin Wischmeyer, Erhard PTH1R Mutants Found in Patients with Primary Failure of Tooth Eruption Disrupt G-Protein Signaling |
title | PTH1R Mutants Found in Patients with Primary Failure of Tooth Eruption Disrupt G-Protein Signaling |
title_full | PTH1R Mutants Found in Patients with Primary Failure of Tooth Eruption Disrupt G-Protein Signaling |
title_fullStr | PTH1R Mutants Found in Patients with Primary Failure of Tooth Eruption Disrupt G-Protein Signaling |
title_full_unstemmed | PTH1R Mutants Found in Patients with Primary Failure of Tooth Eruption Disrupt G-Protein Signaling |
title_short | PTH1R Mutants Found in Patients with Primary Failure of Tooth Eruption Disrupt G-Protein Signaling |
title_sort | pth1r mutants found in patients with primary failure of tooth eruption disrupt g-protein signaling |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5127517/ https://www.ncbi.nlm.nih.gov/pubmed/27898723 http://dx.doi.org/10.1371/journal.pone.0167033 |
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