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b3galt6 Knock-Out Zebrafish Recapitulate β3GalT6-Deficiency Disorders in Human and Reveal a Trisaccharide Proteoglycan Linkage Region
Proteoglycans are structurally and functionally diverse biomacromolecules found abundantly on cell membranes and in the extracellular matrix. They consist of a core protein linked to glycosaminoglycan chains via a tetrasaccharide linkage region. Here, we show that CRISPR/Cas9-mediated b3galt6 knock-...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7758351/ https://www.ncbi.nlm.nih.gov/pubmed/33363150 http://dx.doi.org/10.3389/fcell.2020.597857 |
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author | Delbaere, Sarah De Clercq, Adelbert Mizumoto, Shuji Noborn, Fredrik Bek, Jan Willem Alluyn, Lien Gistelinck, Charlotte Syx, Delfien Salmon, Phil L. Coucke, Paul J. Larson, Göran Yamada, Shuhei Willaert, Andy Malfait, Fransiska |
author_facet | Delbaere, Sarah De Clercq, Adelbert Mizumoto, Shuji Noborn, Fredrik Bek, Jan Willem Alluyn, Lien Gistelinck, Charlotte Syx, Delfien Salmon, Phil L. Coucke, Paul J. Larson, Göran Yamada, Shuhei Willaert, Andy Malfait, Fransiska |
author_sort | Delbaere, Sarah |
collection | PubMed |
description | Proteoglycans are structurally and functionally diverse biomacromolecules found abundantly on cell membranes and in the extracellular matrix. They consist of a core protein linked to glycosaminoglycan chains via a tetrasaccharide linkage region. Here, we show that CRISPR/Cas9-mediated b3galt6 knock-out zebrafish, lacking galactosyltransferase II, which adds the third sugar in the linkage region, largely recapitulate the phenotypic abnormalities seen in human β3GalT6-deficiency disorders. These comprise craniofacial dysmorphism, generalized skeletal dysplasia, skin involvement and indications for muscle hypotonia. In-depth TEM analysis revealed disturbed collagen fibril organization as the most consistent ultrastructural characteristic throughout different affected tissues. Strikingly, despite a strong reduction in glycosaminoglycan content, as demonstrated by anion-exchange HPLC, subsequent LC-MS/MS analysis revealed a small amount of proteoglycans containing a unique linkage region consisting of only three sugars. This implies that formation of glycosaminoglycans with an immature linkage region is possible in a pathogenic context. Our study, therefore unveils a novel rescue mechanism for proteoglycan production in the absence of galactosyltransferase II, hereby opening new avenues for therapeutic intervention. |
format | Online Article Text |
id | pubmed-7758351 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-77583512020-12-25 b3galt6 Knock-Out Zebrafish Recapitulate β3GalT6-Deficiency Disorders in Human and Reveal a Trisaccharide Proteoglycan Linkage Region Delbaere, Sarah De Clercq, Adelbert Mizumoto, Shuji Noborn, Fredrik Bek, Jan Willem Alluyn, Lien Gistelinck, Charlotte Syx, Delfien Salmon, Phil L. Coucke, Paul J. Larson, Göran Yamada, Shuhei Willaert, Andy Malfait, Fransiska Front Cell Dev Biol Cell and Developmental Biology Proteoglycans are structurally and functionally diverse biomacromolecules found abundantly on cell membranes and in the extracellular matrix. They consist of a core protein linked to glycosaminoglycan chains via a tetrasaccharide linkage region. Here, we show that CRISPR/Cas9-mediated b3galt6 knock-out zebrafish, lacking galactosyltransferase II, which adds the third sugar in the linkage region, largely recapitulate the phenotypic abnormalities seen in human β3GalT6-deficiency disorders. These comprise craniofacial dysmorphism, generalized skeletal dysplasia, skin involvement and indications for muscle hypotonia. In-depth TEM analysis revealed disturbed collagen fibril organization as the most consistent ultrastructural characteristic throughout different affected tissues. Strikingly, despite a strong reduction in glycosaminoglycan content, as demonstrated by anion-exchange HPLC, subsequent LC-MS/MS analysis revealed a small amount of proteoglycans containing a unique linkage region consisting of only three sugars. This implies that formation of glycosaminoglycans with an immature linkage region is possible in a pathogenic context. Our study, therefore unveils a novel rescue mechanism for proteoglycan production in the absence of galactosyltransferase II, hereby opening new avenues for therapeutic intervention. Frontiers Media S.A. 2020-12-10 /pmc/articles/PMC7758351/ /pubmed/33363150 http://dx.doi.org/10.3389/fcell.2020.597857 Text en Copyright © 2020 Delbaere, De Clercq, Mizumoto, Noborn, Bek, Alluyn, Gistelinck, Syx, Salmon, Coucke, Larson, Yamada, Willaert and Malfait. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Cell and Developmental Biology Delbaere, Sarah De Clercq, Adelbert Mizumoto, Shuji Noborn, Fredrik Bek, Jan Willem Alluyn, Lien Gistelinck, Charlotte Syx, Delfien Salmon, Phil L. Coucke, Paul J. Larson, Göran Yamada, Shuhei Willaert, Andy Malfait, Fransiska b3galt6 Knock-Out Zebrafish Recapitulate β3GalT6-Deficiency Disorders in Human and Reveal a Trisaccharide Proteoglycan Linkage Region |
title | b3galt6 Knock-Out Zebrafish Recapitulate β3GalT6-Deficiency Disorders in Human and Reveal a Trisaccharide Proteoglycan Linkage Region |
title_full | b3galt6 Knock-Out Zebrafish Recapitulate β3GalT6-Deficiency Disorders in Human and Reveal a Trisaccharide Proteoglycan Linkage Region |
title_fullStr | b3galt6 Knock-Out Zebrafish Recapitulate β3GalT6-Deficiency Disorders in Human and Reveal a Trisaccharide Proteoglycan Linkage Region |
title_full_unstemmed | b3galt6 Knock-Out Zebrafish Recapitulate β3GalT6-Deficiency Disorders in Human and Reveal a Trisaccharide Proteoglycan Linkage Region |
title_short | b3galt6 Knock-Out Zebrafish Recapitulate β3GalT6-Deficiency Disorders in Human and Reveal a Trisaccharide Proteoglycan Linkage Region |
title_sort | b3galt6 knock-out zebrafish recapitulate β3galt6-deficiency disorders in human and reveal a trisaccharide proteoglycan linkage region |
topic | Cell and Developmental Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7758351/ https://www.ncbi.nlm.nih.gov/pubmed/33363150 http://dx.doi.org/10.3389/fcell.2020.597857 |
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