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Protein O-Glucosyltransferase 1 (POGLUT1) Promotes Mouse Gastrulation through Modification of the Apical Polarity Protein CRUMBS2

Crumbs family proteins are apical transmembrane proteins with ancient roles in cell polarity. Mouse Crumbs2 mutants arrest at midgestation with abnormal neural plate morphology and a deficit of mesoderm caused by defects in gastrulation. We identified an ENU-induced mutation, wsnp, that phenocopies...

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Autores principales: Ramkumar, Nitya, Harvey, Beth M., Lee, Jeffrey D., Alcorn, Heather L., Silva-Gagliardi, Nancy F., McGlade, C. Jane, Bestor, Timothy H., Wijnholds, Jan, Haltiwanger, Robert S., Anderson, Kathryn V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4619674/
https://www.ncbi.nlm.nih.gov/pubmed/26496195
http://dx.doi.org/10.1371/journal.pgen.1005551
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author Ramkumar, Nitya
Harvey, Beth M.
Lee, Jeffrey D.
Alcorn, Heather L.
Silva-Gagliardi, Nancy F.
McGlade, C. Jane
Bestor, Timothy H.
Wijnholds, Jan
Haltiwanger, Robert S.
Anderson, Kathryn V.
author_facet Ramkumar, Nitya
Harvey, Beth M.
Lee, Jeffrey D.
Alcorn, Heather L.
Silva-Gagliardi, Nancy F.
McGlade, C. Jane
Bestor, Timothy H.
Wijnholds, Jan
Haltiwanger, Robert S.
Anderson, Kathryn V.
author_sort Ramkumar, Nitya
collection PubMed
description Crumbs family proteins are apical transmembrane proteins with ancient roles in cell polarity. Mouse Crumbs2 mutants arrest at midgestation with abnormal neural plate morphology and a deficit of mesoderm caused by defects in gastrulation. We identified an ENU-induced mutation, wsnp, that phenocopies the Crumbs2 null phenotype. We show that wsnp is a null allele of Protein O-glucosyltransferase 1 (Poglut1), which encodes an enzyme previously shown to add O-glucose to EGF repeats in the extracellular domain of Drosophila and mammalian Notch, but the role of POGLUT1 in mammalian gastrulation has not been investigated. As predicted, we find that POGLUT1 is essential for Notch signaling in the early mouse embryo. However, the loss of mouse POGLUT1 causes an earlier and more dramatic phenotype than does the loss of activity of the Notch pathway, indicating that POGLUT1 has additional biologically relevant substrates. Using mass spectrometry, we show that POGLUT1 modifies EGF repeats in the extracellular domain of full-length mouse CRUMBS2. CRUMBS2 that lacks the O-glucose modification fails to be enriched on the apical plasma membrane and instead accumulates in the endoplasmic reticulum. The data demonstrate that CRUMBS2 is the target of POGLUT1 for the gastrulation epithelial-to-mesenchymal transitions (EMT) and that all activity of CRUMBS2 depends on modification by POGLUT1. Mutations in human POGLUT1 cause Dowling-Degos Disease, POGLUT1 is overexpressed in a variety of tumor cells, and mutations in the EGF repeats of human CRUMBS proteins are associated with human congenital nephrosis, retinitis pigmentosa and retinal degeneration, suggesting that O-glucosylation of CRUMBS proteins has broad roles in human health.
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spelling pubmed-46196742015-10-29 Protein O-Glucosyltransferase 1 (POGLUT1) Promotes Mouse Gastrulation through Modification of the Apical Polarity Protein CRUMBS2 Ramkumar, Nitya Harvey, Beth M. Lee, Jeffrey D. Alcorn, Heather L. Silva-Gagliardi, Nancy F. McGlade, C. Jane Bestor, Timothy H. Wijnholds, Jan Haltiwanger, Robert S. Anderson, Kathryn V. PLoS Genet Research Article Crumbs family proteins are apical transmembrane proteins with ancient roles in cell polarity. Mouse Crumbs2 mutants arrest at midgestation with abnormal neural plate morphology and a deficit of mesoderm caused by defects in gastrulation. We identified an ENU-induced mutation, wsnp, that phenocopies the Crumbs2 null phenotype. We show that wsnp is a null allele of Protein O-glucosyltransferase 1 (Poglut1), which encodes an enzyme previously shown to add O-glucose to EGF repeats in the extracellular domain of Drosophila and mammalian Notch, but the role of POGLUT1 in mammalian gastrulation has not been investigated. As predicted, we find that POGLUT1 is essential for Notch signaling in the early mouse embryo. However, the loss of mouse POGLUT1 causes an earlier and more dramatic phenotype than does the loss of activity of the Notch pathway, indicating that POGLUT1 has additional biologically relevant substrates. Using mass spectrometry, we show that POGLUT1 modifies EGF repeats in the extracellular domain of full-length mouse CRUMBS2. CRUMBS2 that lacks the O-glucose modification fails to be enriched on the apical plasma membrane and instead accumulates in the endoplasmic reticulum. The data demonstrate that CRUMBS2 is the target of POGLUT1 for the gastrulation epithelial-to-mesenchymal transitions (EMT) and that all activity of CRUMBS2 depends on modification by POGLUT1. Mutations in human POGLUT1 cause Dowling-Degos Disease, POGLUT1 is overexpressed in a variety of tumor cells, and mutations in the EGF repeats of human CRUMBS proteins are associated with human congenital nephrosis, retinitis pigmentosa and retinal degeneration, suggesting that O-glucosylation of CRUMBS proteins has broad roles in human health. Public Library of Science 2015-10-23 /pmc/articles/PMC4619674/ /pubmed/26496195 http://dx.doi.org/10.1371/journal.pgen.1005551 Text en © 2015 Ramkumar 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
Ramkumar, Nitya
Harvey, Beth M.
Lee, Jeffrey D.
Alcorn, Heather L.
Silva-Gagliardi, Nancy F.
McGlade, C. Jane
Bestor, Timothy H.
Wijnholds, Jan
Haltiwanger, Robert S.
Anderson, Kathryn V.
Protein O-Glucosyltransferase 1 (POGLUT1) Promotes Mouse Gastrulation through Modification of the Apical Polarity Protein CRUMBS2
title Protein O-Glucosyltransferase 1 (POGLUT1) Promotes Mouse Gastrulation through Modification of the Apical Polarity Protein CRUMBS2
title_full Protein O-Glucosyltransferase 1 (POGLUT1) Promotes Mouse Gastrulation through Modification of the Apical Polarity Protein CRUMBS2
title_fullStr Protein O-Glucosyltransferase 1 (POGLUT1) Promotes Mouse Gastrulation through Modification of the Apical Polarity Protein CRUMBS2
title_full_unstemmed Protein O-Glucosyltransferase 1 (POGLUT1) Promotes Mouse Gastrulation through Modification of the Apical Polarity Protein CRUMBS2
title_short Protein O-Glucosyltransferase 1 (POGLUT1) Promotes Mouse Gastrulation through Modification of the Apical Polarity Protein CRUMBS2
title_sort protein o-glucosyltransferase 1 (poglut1) promotes mouse gastrulation through modification of the apical polarity protein crumbs2
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4619674/
https://www.ncbi.nlm.nih.gov/pubmed/26496195
http://dx.doi.org/10.1371/journal.pgen.1005551
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