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Characterization of Novel MSX1 Mutations Identified in Japanese Patients with Nonsyndromic Tooth Agenesis

Since MSX1 and PAX9 are linked to the pathogenesis of nonsyndromic tooth agenesis, we performed detailed mutational analysis of these two genes sampled from Japanese patients. We identified two novel MSX1 variants with an amino acid substitution within the homeodomain; Thr174Ile (T174I) from a spora...

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Autores principales: Yamaguchi, Seishi, Machida, Junichiro, Kamamoto, Munefumi, Kimura, Masashi, Shibata, Akio, Tatematsu, Tadashi, Miyachi, Hitoshi, Higashi, Yujiro, Jezewski, Peter, Nakayama, Atsuo, Shimozato, Kazuo, Tokita, Yoshihito
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4125152/
https://www.ncbi.nlm.nih.gov/pubmed/25101640
http://dx.doi.org/10.1371/journal.pone.0102944
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author Yamaguchi, Seishi
Machida, Junichiro
Kamamoto, Munefumi
Kimura, Masashi
Shibata, Akio
Tatematsu, Tadashi
Miyachi, Hitoshi
Higashi, Yujiro
Jezewski, Peter
Nakayama, Atsuo
Shimozato, Kazuo
Tokita, Yoshihito
author_facet Yamaguchi, Seishi
Machida, Junichiro
Kamamoto, Munefumi
Kimura, Masashi
Shibata, Akio
Tatematsu, Tadashi
Miyachi, Hitoshi
Higashi, Yujiro
Jezewski, Peter
Nakayama, Atsuo
Shimozato, Kazuo
Tokita, Yoshihito
author_sort Yamaguchi, Seishi
collection PubMed
description Since MSX1 and PAX9 are linked to the pathogenesis of nonsyndromic tooth agenesis, we performed detailed mutational analysis of these two genes sampled from Japanese patients. We identified two novel MSX1 variants with an amino acid substitution within the homeodomain; Thr174Ile (T174I) from a sporadic hypodontia case and Leu205Arg (L205R) from a familial oligodontia case. Both the Thr174 and Leu205 residues in the MSX1 homeodomain are highly conserved among different species. To define possible roles of mutations at these amino acids in the pathogenesis of nonsyndromic tooth agenesis, we performed several functional analyses. It has been demonstrated that MSX1 plays a pivotal role in hard tissue development as a suppressor for mesenchymal cell differentiation. To evaluate the suppression activity of the variants in mesenchymal cells, we used the myoD-promoter, which is one of convenient reporter assay system for MSX1. Although the gene products of these MSX1 variants are stable and capable of normal nuclear localization, they do not suppress myoD-promoter activity in differentiated C2C12 cells. To clarify the molecular mechanisms underlying our results, we performed further analyses including electrophoretic mobility shift assays, and co-immunoprecipitation assays to survey the molecular interactions between the mutant MSX1 proteins and the oligonucleotide DNA with MSX1 consensus binding motif or EZH2 methyltransferase. Since EZH2 is reported to interact with MSX1 and regulate MSX1 mediated gene suppression, we hypothesized that the T174I and L205R substitutions would impair this interaction. We conclude from the results of our experiments that the DNA binding ability of MSX1 is abolished by these two amino acid substitutions. This illustrates a causative role of the T174I and L205R MSX1 homeodomain mutations in tooth agenesis, and suggests that they may influence cell proliferation and differentiation resulting in lesser tooth germ formation in vivo.
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spelling pubmed-41251522014-08-12 Characterization of Novel MSX1 Mutations Identified in Japanese Patients with Nonsyndromic Tooth Agenesis Yamaguchi, Seishi Machida, Junichiro Kamamoto, Munefumi Kimura, Masashi Shibata, Akio Tatematsu, Tadashi Miyachi, Hitoshi Higashi, Yujiro Jezewski, Peter Nakayama, Atsuo Shimozato, Kazuo Tokita, Yoshihito PLoS One Research Article Since MSX1 and PAX9 are linked to the pathogenesis of nonsyndromic tooth agenesis, we performed detailed mutational analysis of these two genes sampled from Japanese patients. We identified two novel MSX1 variants with an amino acid substitution within the homeodomain; Thr174Ile (T174I) from a sporadic hypodontia case and Leu205Arg (L205R) from a familial oligodontia case. Both the Thr174 and Leu205 residues in the MSX1 homeodomain are highly conserved among different species. To define possible roles of mutations at these amino acids in the pathogenesis of nonsyndromic tooth agenesis, we performed several functional analyses. It has been demonstrated that MSX1 plays a pivotal role in hard tissue development as a suppressor for mesenchymal cell differentiation. To evaluate the suppression activity of the variants in mesenchymal cells, we used the myoD-promoter, which is one of convenient reporter assay system for MSX1. Although the gene products of these MSX1 variants are stable and capable of normal nuclear localization, they do not suppress myoD-promoter activity in differentiated C2C12 cells. To clarify the molecular mechanisms underlying our results, we performed further analyses including electrophoretic mobility shift assays, and co-immunoprecipitation assays to survey the molecular interactions between the mutant MSX1 proteins and the oligonucleotide DNA with MSX1 consensus binding motif or EZH2 methyltransferase. Since EZH2 is reported to interact with MSX1 and regulate MSX1 mediated gene suppression, we hypothesized that the T174I and L205R substitutions would impair this interaction. We conclude from the results of our experiments that the DNA binding ability of MSX1 is abolished by these two amino acid substitutions. This illustrates a causative role of the T174I and L205R MSX1 homeodomain mutations in tooth agenesis, and suggests that they may influence cell proliferation and differentiation resulting in lesser tooth germ formation in vivo. Public Library of Science 2014-08-07 /pmc/articles/PMC4125152/ /pubmed/25101640 http://dx.doi.org/10.1371/journal.pone.0102944 Text en © 2014 Yamaguchi 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, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Yamaguchi, Seishi
Machida, Junichiro
Kamamoto, Munefumi
Kimura, Masashi
Shibata, Akio
Tatematsu, Tadashi
Miyachi, Hitoshi
Higashi, Yujiro
Jezewski, Peter
Nakayama, Atsuo
Shimozato, Kazuo
Tokita, Yoshihito
Characterization of Novel MSX1 Mutations Identified in Japanese Patients with Nonsyndromic Tooth Agenesis
title Characterization of Novel MSX1 Mutations Identified in Japanese Patients with Nonsyndromic Tooth Agenesis
title_full Characterization of Novel MSX1 Mutations Identified in Japanese Patients with Nonsyndromic Tooth Agenesis
title_fullStr Characterization of Novel MSX1 Mutations Identified in Japanese Patients with Nonsyndromic Tooth Agenesis
title_full_unstemmed Characterization of Novel MSX1 Mutations Identified in Japanese Patients with Nonsyndromic Tooth Agenesis
title_short Characterization of Novel MSX1 Mutations Identified in Japanese Patients with Nonsyndromic Tooth Agenesis
title_sort characterization of novel msx1 mutations identified in japanese patients with nonsyndromic tooth agenesis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4125152/
https://www.ncbi.nlm.nih.gov/pubmed/25101640
http://dx.doi.org/10.1371/journal.pone.0102944
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