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Golgi targeting of Drosophila melanogaster β4GalNAcTB requires a DHHC protein family–related protein as a pilot

Drosophila melanogaster β4GalNAcTB mutant flies revealed that this particular N-acetylgalactosaminyltransferase is predominant in the formation of lacdiNAc (GalNAcβ1,4GlcNAc)-modified glycolipids, but enzymatic activity could not be confirmed for the cloned enzyme. Using a heterologous expression cl...

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Detalles Bibliográficos
Autores principales: Johswich, Anita, Kraft, Benjamin, Wuhrer, Manfred, Berger, Monika, Deelder, André M., Hokke, Cornelis H., Gerardy-Schahn, Rita, Bakker, Hans
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2615082/
https://www.ncbi.nlm.nih.gov/pubmed/19139268
http://dx.doi.org/10.1083/jcb.200801071
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author Johswich, Anita
Kraft, Benjamin
Wuhrer, Manfred
Berger, Monika
Deelder, André M.
Hokke, Cornelis H.
Gerardy-Schahn, Rita
Bakker, Hans
author_facet Johswich, Anita
Kraft, Benjamin
Wuhrer, Manfred
Berger, Monika
Deelder, André M.
Hokke, Cornelis H.
Gerardy-Schahn, Rita
Bakker, Hans
author_sort Johswich, Anita
collection PubMed
description Drosophila melanogaster β4GalNAcTB mutant flies revealed that this particular N-acetylgalactosaminyltransferase is predominant in the formation of lacdiNAc (GalNAcβ1,4GlcNAc)-modified glycolipids, but enzymatic activity could not be confirmed for the cloned enzyme. Using a heterologous expression cloning approach, we isolated β4GalNAcTB together with β4GalNAcTB pilot (GABPI), a multimembrane-spanning protein related to Asp-His-His-Cys (DHHC) proteins but lacking the DHHC consensus sequence. In the absence of GABPI, inactive β4GalNAcTB is trapped in the endoplasmic reticulum (ER). Coexpression of β4GalNAcTB and GABPI generates the active enzyme that is localized together with GABPI in the Golgi. GABPI associates with β4GalNAcTB and, when expressed with an ER retention signal, holds active β4GalNAcTB in the ER. Importantly, treatment of isolated membrane vesicles with Triton X-100 disturbs β4GalNAcTB activity. This phenomenon occurs with multimembrane-spanning glycosyltransferases but is normally not a property of glycosyltransferases with one membrane anchor. In summary, our data provide evidence that GABPI is required for ER export and activity of β4GalNAcTB.
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spelling pubmed-26150822009-07-12 Golgi targeting of Drosophila melanogaster β4GalNAcTB requires a DHHC protein family–related protein as a pilot Johswich, Anita Kraft, Benjamin Wuhrer, Manfred Berger, Monika Deelder, André M. Hokke, Cornelis H. Gerardy-Schahn, Rita Bakker, Hans J Cell Biol Research Articles Drosophila melanogaster β4GalNAcTB mutant flies revealed that this particular N-acetylgalactosaminyltransferase is predominant in the formation of lacdiNAc (GalNAcβ1,4GlcNAc)-modified glycolipids, but enzymatic activity could not be confirmed for the cloned enzyme. Using a heterologous expression cloning approach, we isolated β4GalNAcTB together with β4GalNAcTB pilot (GABPI), a multimembrane-spanning protein related to Asp-His-His-Cys (DHHC) proteins but lacking the DHHC consensus sequence. In the absence of GABPI, inactive β4GalNAcTB is trapped in the endoplasmic reticulum (ER). Coexpression of β4GalNAcTB and GABPI generates the active enzyme that is localized together with GABPI in the Golgi. GABPI associates with β4GalNAcTB and, when expressed with an ER retention signal, holds active β4GalNAcTB in the ER. Importantly, treatment of isolated membrane vesicles with Triton X-100 disturbs β4GalNAcTB activity. This phenomenon occurs with multimembrane-spanning glycosyltransferases but is normally not a property of glycosyltransferases with one membrane anchor. In summary, our data provide evidence that GABPI is required for ER export and activity of β4GalNAcTB. The Rockefeller University Press 2009-01-12 /pmc/articles/PMC2615082/ /pubmed/19139268 http://dx.doi.org/10.1083/jcb.200801071 Text en © 2009 Johswich et al. This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).
spellingShingle Research Articles
Johswich, Anita
Kraft, Benjamin
Wuhrer, Manfred
Berger, Monika
Deelder, André M.
Hokke, Cornelis H.
Gerardy-Schahn, Rita
Bakker, Hans
Golgi targeting of Drosophila melanogaster β4GalNAcTB requires a DHHC protein family–related protein as a pilot
title Golgi targeting of Drosophila melanogaster β4GalNAcTB requires a DHHC protein family–related protein as a pilot
title_full Golgi targeting of Drosophila melanogaster β4GalNAcTB requires a DHHC protein family–related protein as a pilot
title_fullStr Golgi targeting of Drosophila melanogaster β4GalNAcTB requires a DHHC protein family–related protein as a pilot
title_full_unstemmed Golgi targeting of Drosophila melanogaster β4GalNAcTB requires a DHHC protein family–related protein as a pilot
title_short Golgi targeting of Drosophila melanogaster β4GalNAcTB requires a DHHC protein family–related protein as a pilot
title_sort golgi targeting of drosophila melanogaster β4galnactb requires a dhhc protein family–related protein as a pilot
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2615082/
https://www.ncbi.nlm.nih.gov/pubmed/19139268
http://dx.doi.org/10.1083/jcb.200801071
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