Heparan sulfate proteoglycans serve as alternative receptors for low affinity LCMV variants.

Members of the Old World Arenaviruses primarily utilize α-dystroglycan (α-DAG1) as a cellular receptor for infection. Mutations within the glycoprotein (GP) of lymphocytic choriomeningitis virus (LCMV) reduce or abrogate the binding affinity to α-DAG1 and thus influence viral persistence, kinetics,...

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Autores principales:	André Volland, Michael Lohmüller, Emmanuel Heilmann, Janine Kimpel, Sebastian Herzog, Dorothee von Laer
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/71ffc4d83c2e4206961d501abc460c3d
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spelling	oai:doaj.org-article:71ffc4d83c2e4206961d501abc460c3d2021-12-02T20:00:01ZHeparan sulfate proteoglycans serve as alternative receptors for low affinity LCMV variants.1553-73661553-737410.1371/journal.ppat.1009996https://doaj.org/article/71ffc4d83c2e4206961d501abc460c3d2021-10-01T00:00:00Zhttps://doi.org/10.1371/journal.ppat.1009996https://doaj.org/toc/1553-7366https://doaj.org/toc/1553-7374Members of the Old World Arenaviruses primarily utilize α-dystroglycan (α-DAG1) as a cellular receptor for infection. Mutations within the glycoprotein (GP) of lymphocytic choriomeningitis virus (LCMV) reduce or abrogate the binding affinity to α-DAG1 and thus influence viral persistence, kinetics, and cell tropism. The observation that α-DAG1 deficient cells are still highly susceptible to low affinity variants, suggests the use of an alternative receptor(s). In this study, we used a genome-wide CRISPR Cas9 knockout screen in DAG1 deficient 293T cells to identify host factors involved in α-DAG1-independent LCMV infection. By challenging cells with vesicular stomatitis virus (VSV), pseudotyped with the GP of LCMV WE HPI (VSV-GP), we identified the heparan sulfate (HS) biosynthesis pathway as an important host factor for low affinity LCMV infection. These results were confirmed by a genetic approach targeting EXTL3, a key factor in the HS biosynthesis pathway, as well as by enzymatic and chemical methods. Interestingly, a single point mutation within GP1 (S153F or Y155H) of WE HPI is sufficient for the switch from DAG1 to HS binding. Furthermore, we established a simple and reliable virus-binding assay, using directly labelled VSV-GP by intramolecular fusion of VSV-P and mWasabi, demonstrating the importance of HS for virus attachment but not entry in Burkitt lymphoma cells after reconstitution of HS expression. Collectively, our study highlights the essential role of HS for low affinity LCMV infection in contrast to their high affinity counterparts. Residual LCMV infection in double knockouts indicate the use of (a) still unknown entry receptor(s).André VollandMichael LohmüllerEmmanuel HeilmannJanine KimpelSebastian HerzogDorothee von LaerPublic Library of Science (PLoS)articleImmunologic diseases. AllergyRC581-607Biology (General)QH301-705.5ENPLoS Pathogens, Vol 17, Iss 10, p e1009996 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5
spellingShingle	Immunologic diseases. Allergy RC581-607 Biology (General) QH301-705.5 André Volland Michael Lohmüller Emmanuel Heilmann Janine Kimpel Sebastian Herzog Dorothee von Laer Heparan sulfate proteoglycans serve as alternative receptors for low affinity LCMV variants.
description	Members of the Old World Arenaviruses primarily utilize α-dystroglycan (α-DAG1) as a cellular receptor for infection. Mutations within the glycoprotein (GP) of lymphocytic choriomeningitis virus (LCMV) reduce or abrogate the binding affinity to α-DAG1 and thus influence viral persistence, kinetics, and cell tropism. The observation that α-DAG1 deficient cells are still highly susceptible to low affinity variants, suggests the use of an alternative receptor(s). In this study, we used a genome-wide CRISPR Cas9 knockout screen in DAG1 deficient 293T cells to identify host factors involved in α-DAG1-independent LCMV infection. By challenging cells with vesicular stomatitis virus (VSV), pseudotyped with the GP of LCMV WE HPI (VSV-GP), we identified the heparan sulfate (HS) biosynthesis pathway as an important host factor for low affinity LCMV infection. These results were confirmed by a genetic approach targeting EXTL3, a key factor in the HS biosynthesis pathway, as well as by enzymatic and chemical methods. Interestingly, a single point mutation within GP1 (S153F or Y155H) of WE HPI is sufficient for the switch from DAG1 to HS binding. Furthermore, we established a simple and reliable virus-binding assay, using directly labelled VSV-GP by intramolecular fusion of VSV-P and mWasabi, demonstrating the importance of HS for virus attachment but not entry in Burkitt lymphoma cells after reconstitution of HS expression. Collectively, our study highlights the essential role of HS for low affinity LCMV infection in contrast to their high affinity counterparts. Residual LCMV infection in double knockouts indicate the use of (a) still unknown entry receptor(s).
format	article
author	André Volland Michael Lohmüller Emmanuel Heilmann Janine Kimpel Sebastian Herzog Dorothee von Laer
author_facet	André Volland Michael Lohmüller Emmanuel Heilmann Janine Kimpel Sebastian Herzog Dorothee von Laer
author_sort	André Volland
title	Heparan sulfate proteoglycans serve as alternative receptors for low affinity LCMV variants.
title_short	Heparan sulfate proteoglycans serve as alternative receptors for low affinity LCMV variants.
title_full	Heparan sulfate proteoglycans serve as alternative receptors for low affinity LCMV variants.
title_fullStr	Heparan sulfate proteoglycans serve as alternative receptors for low affinity LCMV variants.
title_full_unstemmed	Heparan sulfate proteoglycans serve as alternative receptors for low affinity LCMV variants.
title_sort	heparan sulfate proteoglycans serve as alternative receptors for low affinity lcmv variants.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/71ffc4d83c2e4206961d501abc460c3d
work_keys_str_mv	AT andrevolland heparansulfateproteoglycansserveasalternativereceptorsforlowaffinitylcmvvariants AT michaellohmuller heparansulfateproteoglycansserveasalternativereceptorsforlowaffinitylcmvvariants AT emmanuelheilmann heparansulfateproteoglycansserveasalternativereceptorsforlowaffinitylcmvvariants AT janinekimpel heparansulfateproteoglycansserveasalternativereceptorsforlowaffinitylcmvvariants AT sebastianherzog heparansulfateproteoglycansserveasalternativereceptorsforlowaffinitylcmvvariants AT dorotheevonlaer heparansulfateproteoglycansserveasalternativereceptorsforlowaffinitylcmvvariants
_version_	1718375728951590912

Heparan sulfate proteoglycans serve as alternative receptors for low affinity LCMV variants.

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