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The role of N-linked glycosylation in proteolytic processing and cell surface transport of the Cedar virus fusion protein
BACKGROUND: N-linked glycans on viral glycoproteins have been shown to be important for protein expression, processing and intracellular transport. The fusion glycoprotein F of Cedar virus (CedV) contains six potential N-glycosylation sites. FINDINGS: To investigate their impact on cell surface tran...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9400332/ https://www.ncbi.nlm.nih.gov/pubmed/35999637 http://dx.doi.org/10.1186/s12985-022-01864-5 |
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author | Fischer, Kerstin Topallar, Selin Kraatz, Franziska Groschup, Martin H. Diederich, Sandra |
author_facet | Fischer, Kerstin Topallar, Selin Kraatz, Franziska Groschup, Martin H. Diederich, Sandra |
author_sort | Fischer, Kerstin |
collection | PubMed |
description | BACKGROUND: N-linked glycans on viral glycoproteins have been shown to be important for protein expression, processing and intracellular transport. The fusion glycoprotein F of Cedar virus (CedV) contains six potential N-glycosylation sites. FINDINGS: To investigate their impact on cell surface transport, proteolytic cleavage and biological activity, we disrupted the consensus sequences by conservative mutations (Asn to Gln) and found that five of the six potential N-glycosylation sites are actually utilized. The individual removal of N-glycan g1 (N66), g2 (N79) and g3 (N98) in the CedV F(2) subunit had no or only little effect on cell surface transport, proteolytic cleavage and fusion activity of CedV F. Interestingly, removal of N-linked glycan g6 (N463) in the F(1) subunit resulted in reduced cell surface expression but slightly increased fusogenicity upon co-expression with the CedV receptor-binding protein G. Most prominent effects however were observed for the disruption of N-glycosylation motif g4 (N413), which significantly impaired the transport of CedV F to the cell surface, thereby also affecting proteolytic cleavage and fusion activity. CONCLUSIONS: Our findings indicate that the individual N-linked modifications, with the exception of glycan g4, are dispensable for processing of CedV F protein in transfection experiments. However, removal of g4 led to a phenotype that was strongly impaired concerning cell surface expression and proteolytic activation. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12985-022-01864-5. |
format | Online Article Text |
id | pubmed-9400332 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-94003322022-08-25 The role of N-linked glycosylation in proteolytic processing and cell surface transport of the Cedar virus fusion protein Fischer, Kerstin Topallar, Selin Kraatz, Franziska Groschup, Martin H. Diederich, Sandra Virol J Research BACKGROUND: N-linked glycans on viral glycoproteins have been shown to be important for protein expression, processing and intracellular transport. The fusion glycoprotein F of Cedar virus (CedV) contains six potential N-glycosylation sites. FINDINGS: To investigate their impact on cell surface transport, proteolytic cleavage and biological activity, we disrupted the consensus sequences by conservative mutations (Asn to Gln) and found that five of the six potential N-glycosylation sites are actually utilized. The individual removal of N-glycan g1 (N66), g2 (N79) and g3 (N98) in the CedV F(2) subunit had no or only little effect on cell surface transport, proteolytic cleavage and fusion activity of CedV F. Interestingly, removal of N-linked glycan g6 (N463) in the F(1) subunit resulted in reduced cell surface expression but slightly increased fusogenicity upon co-expression with the CedV receptor-binding protein G. Most prominent effects however were observed for the disruption of N-glycosylation motif g4 (N413), which significantly impaired the transport of CedV F to the cell surface, thereby also affecting proteolytic cleavage and fusion activity. CONCLUSIONS: Our findings indicate that the individual N-linked modifications, with the exception of glycan g4, are dispensable for processing of CedV F protein in transfection experiments. However, removal of g4 led to a phenotype that was strongly impaired concerning cell surface expression and proteolytic activation. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12985-022-01864-5. BioMed Central 2022-08-23 /pmc/articles/PMC9400332/ /pubmed/35999637 http://dx.doi.org/10.1186/s12985-022-01864-5 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Fischer, Kerstin Topallar, Selin Kraatz, Franziska Groschup, Martin H. Diederich, Sandra The role of N-linked glycosylation in proteolytic processing and cell surface transport of the Cedar virus fusion protein |
title | The role of N-linked glycosylation in proteolytic processing and cell surface transport of the Cedar virus fusion protein |
title_full | The role of N-linked glycosylation in proteolytic processing and cell surface transport of the Cedar virus fusion protein |
title_fullStr | The role of N-linked glycosylation in proteolytic processing and cell surface transport of the Cedar virus fusion protein |
title_full_unstemmed | The role of N-linked glycosylation in proteolytic processing and cell surface transport of the Cedar virus fusion protein |
title_short | The role of N-linked glycosylation in proteolytic processing and cell surface transport of the Cedar virus fusion protein |
title_sort | role of n-linked glycosylation in proteolytic processing and cell surface transport of the cedar virus fusion protein |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9400332/ https://www.ncbi.nlm.nih.gov/pubmed/35999637 http://dx.doi.org/10.1186/s12985-022-01864-5 |
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