Dissecting virus entry: replication-independent analysis of virus binding, internalization, and penetration using minimal complementation of β-galactosidase.

Studies of viral entry into host cells often rely on the detection of post-entry parameters, such as viral replication or the expression of a reporter gene, rather than on measuring entry per se. The lack of assays to easily detect the different steps of entry severely hampers the analysis of this k...

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Autores principales:	Christine Burkard, Louis-Marie Bloyet, Oliver Wicht, Frank J van Kuppeveld, Peter J M Rottier, Cornelis A M de Haan, Berend Jan Bosch
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/39de291686d94fdaabb6cf7a2379338a
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id	oai:doaj.org-article:39de291686d94fdaabb6cf7a2379338a
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spelling	oai:doaj.org-article:39de291686d94fdaabb6cf7a2379338a2021-11-25T06:08:28ZDissecting virus entry: replication-independent analysis of virus binding, internalization, and penetration using minimal complementation of β-galactosidase.1932-620310.1371/journal.pone.0101762https://doaj.org/article/39de291686d94fdaabb6cf7a2379338a2014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/25025332/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Studies of viral entry into host cells often rely on the detection of post-entry parameters, such as viral replication or the expression of a reporter gene, rather than on measuring entry per se. The lack of assays to easily detect the different steps of entry severely hampers the analysis of this key process in virus infection. Here we describe novel, highly adaptable viral entry assays making use of minimal complementation of the E. coli β-galactosidase in mammalian cells. Enzyme activity is reconstituted when a small intravirion peptide (α-peptide) is complementing the inactive mutant form ΔM15 of β-galactosidase. The method allows to dissect and to independently detect binding, internalization, and fusion of viruses during host cell entry. Here we use it to confirm and extend current knowledge on the entry process of two enveloped viruses: vesicular stomatitis virus (VSV) and murine hepatitis coronavirus (MHV).Christine BurkardLouis-Marie BloyetOliver WichtFrank J van KuppeveldPeter J M RottierCornelis A M de HaanBerend Jan BoschPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 7, p e101762 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Christine Burkard Louis-Marie Bloyet Oliver Wicht Frank J van Kuppeveld Peter J M Rottier Cornelis A M de Haan Berend Jan Bosch Dissecting virus entry: replication-independent analysis of virus binding, internalization, and penetration using minimal complementation of β-galactosidase.
description	Studies of viral entry into host cells often rely on the detection of post-entry parameters, such as viral replication or the expression of a reporter gene, rather than on measuring entry per se. The lack of assays to easily detect the different steps of entry severely hampers the analysis of this key process in virus infection. Here we describe novel, highly adaptable viral entry assays making use of minimal complementation of the E. coli β-galactosidase in mammalian cells. Enzyme activity is reconstituted when a small intravirion peptide (α-peptide) is complementing the inactive mutant form ΔM15 of β-galactosidase. The method allows to dissect and to independently detect binding, internalization, and fusion of viruses during host cell entry. Here we use it to confirm and extend current knowledge on the entry process of two enveloped viruses: vesicular stomatitis virus (VSV) and murine hepatitis coronavirus (MHV).
format	article
author	Christine Burkard Louis-Marie Bloyet Oliver Wicht Frank J van Kuppeveld Peter J M Rottier Cornelis A M de Haan Berend Jan Bosch
author_facet	Christine Burkard Louis-Marie Bloyet Oliver Wicht Frank J van Kuppeveld Peter J M Rottier Cornelis A M de Haan Berend Jan Bosch
author_sort	Christine Burkard
title	Dissecting virus entry: replication-independent analysis of virus binding, internalization, and penetration using minimal complementation of β-galactosidase.
title_short	Dissecting virus entry: replication-independent analysis of virus binding, internalization, and penetration using minimal complementation of β-galactosidase.
title_full	Dissecting virus entry: replication-independent analysis of virus binding, internalization, and penetration using minimal complementation of β-galactosidase.
title_fullStr	Dissecting virus entry: replication-independent analysis of virus binding, internalization, and penetration using minimal complementation of β-galactosidase.
title_full_unstemmed	Dissecting virus entry: replication-independent analysis of virus binding, internalization, and penetration using minimal complementation of β-galactosidase.
title_sort	dissecting virus entry: replication-independent analysis of virus binding, internalization, and penetration using minimal complementation of β-galactosidase.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/39de291686d94fdaabb6cf7a2379338a
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Dissecting virus entry: replication-independent analysis of virus binding, internalization, and penetration using minimal complementation of β-galactosidase.

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