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N-glycosylation in the Pre-Membrane Protein Is Essential for the Zika Virus Life Cycle

Asparagine (N)-linked protein glycosylation plays an important role in protein synthesis and modification. Two Zika virus (ZIKV) structural proteins, the pre-membrane (prM) and envelope (E) protein are N-glycosylated. The prM protein of all ZIKV strains contains a single N-linked glycosylation site,...

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Autores principales: Gwon, Yong-Dae, Zusinaite, Eva, Merits, Andres, Överby, Anna K., Evander, Magnus
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7552079/
https://www.ncbi.nlm.nih.gov/pubmed/32842538
http://dx.doi.org/10.3390/v12090925
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author Gwon, Yong-Dae
Zusinaite, Eva
Merits, Andres
Överby, Anna K.
Evander, Magnus
author_facet Gwon, Yong-Dae
Zusinaite, Eva
Merits, Andres
Överby, Anna K.
Evander, Magnus
author_sort Gwon, Yong-Dae
collection PubMed
description Asparagine (N)-linked protein glycosylation plays an important role in protein synthesis and modification. Two Zika virus (ZIKV) structural proteins, the pre-membrane (prM) and envelope (E) protein are N-glycosylated. The prM protein of all ZIKV strains contains a single N-linked glycosylation site, while not all strains contain an N-linked site in the E protein. Our aim was to examine the impact of prM and E N-linked glycosylation on ZIKV infectivity and cell trafficking. Using a ZIKV infectious clone, we found that when the N-glycan sites were removed, the prM- and the prM/E-double mutants did not produce an infectious virus in the supernatant. Further, by using ZIKV prME constructs, we found that N-glycosylation was necessary for effective secretion of ZIKV virions. The absence of the N-glycan on prM or E caused protein aggregation in the rough endoplasmatic reticulum (ER) compartment. The aggregation was more pronounced for the prM-mutation, and the mutant virus lost the ER-Golgi intermediate compartment (ERGIC) localization. In addition, lack of the N-glycan on prM induced nuclear translocation of CCAAT-enhancer-binding protein homologous protein (CHOP), an ER stress marker. To conclude, we show that the prM N-glycan is essential for the ZIKV infectious cycle, and plays an important role in viral protein trafficking, protein folding, and virion assembly.
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spelling pubmed-75520792020-10-14 N-glycosylation in the Pre-Membrane Protein Is Essential for the Zika Virus Life Cycle Gwon, Yong-Dae Zusinaite, Eva Merits, Andres Överby, Anna K. Evander, Magnus Viruses Article Asparagine (N)-linked protein glycosylation plays an important role in protein synthesis and modification. Two Zika virus (ZIKV) structural proteins, the pre-membrane (prM) and envelope (E) protein are N-glycosylated. The prM protein of all ZIKV strains contains a single N-linked glycosylation site, while not all strains contain an N-linked site in the E protein. Our aim was to examine the impact of prM and E N-linked glycosylation on ZIKV infectivity and cell trafficking. Using a ZIKV infectious clone, we found that when the N-glycan sites were removed, the prM- and the prM/E-double mutants did not produce an infectious virus in the supernatant. Further, by using ZIKV prME constructs, we found that N-glycosylation was necessary for effective secretion of ZIKV virions. The absence of the N-glycan on prM or E caused protein aggregation in the rough endoplasmatic reticulum (ER) compartment. The aggregation was more pronounced for the prM-mutation, and the mutant virus lost the ER-Golgi intermediate compartment (ERGIC) localization. In addition, lack of the N-glycan on prM induced nuclear translocation of CCAAT-enhancer-binding protein homologous protein (CHOP), an ER stress marker. To conclude, we show that the prM N-glycan is essential for the ZIKV infectious cycle, and plays an important role in viral protein trafficking, protein folding, and virion assembly. MDPI 2020-08-23 /pmc/articles/PMC7552079/ /pubmed/32842538 http://dx.doi.org/10.3390/v12090925 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Gwon, Yong-Dae
Zusinaite, Eva
Merits, Andres
Överby, Anna K.
Evander, Magnus
N-glycosylation in the Pre-Membrane Protein Is Essential for the Zika Virus Life Cycle
title N-glycosylation in the Pre-Membrane Protein Is Essential for the Zika Virus Life Cycle
title_full N-glycosylation in the Pre-Membrane Protein Is Essential for the Zika Virus Life Cycle
title_fullStr N-glycosylation in the Pre-Membrane Protein Is Essential for the Zika Virus Life Cycle
title_full_unstemmed N-glycosylation in the Pre-Membrane Protein Is Essential for the Zika Virus Life Cycle
title_short N-glycosylation in the Pre-Membrane Protein Is Essential for the Zika Virus Life Cycle
title_sort n-glycosylation in the pre-membrane protein is essential for the zika virus life cycle
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7552079/
https://www.ncbi.nlm.nih.gov/pubmed/32842538
http://dx.doi.org/10.3390/v12090925
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