Influenza A virus encoding secreted Gaussia luciferase as useful tool to analyze viral replication and its inhibition by antiviral compounds and cellular proteins.

Reporter genes inserted into viral genomes enable the easy and rapid quantification of virus replication, which is instrumental to efficient in vitro screening of antiviral compounds or in vivo analysis of viral spread and pathogenesis. Based on a published design, we have generated several replicat...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Nadine Eckert, Florian Wrensch, Sabine Gärtner, Navaneethan Palanisamy, Ulrike Goedecke, Nils Jäger, Stefan Pöhlmann, Michael Winkler
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1c94ba363cd24da1a82b21e43ebe0ccc
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:1c94ba363cd24da1a82b21e43ebe0ccc
record_format	dspace
spelling	oai:doaj.org-article:1c94ba363cd24da1a82b21e43ebe0ccc2021-11-18T08:18:44ZInfluenza A virus encoding secreted Gaussia luciferase as useful tool to analyze viral replication and its inhibition by antiviral compounds and cellular proteins.1932-620310.1371/journal.pone.0097695https://doaj.org/article/1c94ba363cd24da1a82b21e43ebe0ccc2014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24842154/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Reporter genes inserted into viral genomes enable the easy and rapid quantification of virus replication, which is instrumental to efficient in vitro screening of antiviral compounds or in vivo analysis of viral spread and pathogenesis. Based on a published design, we have generated several replication competent influenza A viruses carrying either fluorescent proteins or Gaussia luciferase. Reporter activity could be readily quantified in infected cultures, but the virus encoding Gaussia luciferase was more stable than viruses bearing fluorescent proteins and was therefore analyzed in detail. Quantification of Gaussia luciferase activity in the supernatants of infected culture allowed the convenient and highly sensitive detection of viral spread, and enzymatic activity correlated with the number of infectious particles released from infected cells. Furthermore, the Gaussia luciferase encoding virus allowed the sensitive quantification of the antiviral activity of the neuraminidase inhibitor (NAI) zanamivir and the host cell interferon-inducible transmembrane (IFITM) proteins 1-3, which are known to inhibit influenza virus entry. Finally, the virus was used to demonstrate that influenza A virus infection is sensitive to a modulator of endosomal cholesterol, in keeping with the concept that IFITMs inhibit viral entry by altering cholesterol levels in the endosomal membrane. In sum, we report the characterization of a novel influenza A reporter virus, which allows fast and sensitive detection of viral spread and its inhibition, and we show that influenza A virus entry is sensitive to alterations of endosomal cholesterol levels.Nadine EckertFlorian WrenschSabine GärtnerNavaneethan PalanisamyUlrike GoedeckeNils JägerStefan PöhlmannMichael WinklerPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 5, p e97695 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Nadine Eckert Florian Wrensch Sabine Gärtner Navaneethan Palanisamy Ulrike Goedecke Nils Jäger Stefan Pöhlmann Michael Winkler Influenza A virus encoding secreted Gaussia luciferase as useful tool to analyze viral replication and its inhibition by antiviral compounds and cellular proteins.
description	Reporter genes inserted into viral genomes enable the easy and rapid quantification of virus replication, which is instrumental to efficient in vitro screening of antiviral compounds or in vivo analysis of viral spread and pathogenesis. Based on a published design, we have generated several replication competent influenza A viruses carrying either fluorescent proteins or Gaussia luciferase. Reporter activity could be readily quantified in infected cultures, but the virus encoding Gaussia luciferase was more stable than viruses bearing fluorescent proteins and was therefore analyzed in detail. Quantification of Gaussia luciferase activity in the supernatants of infected culture allowed the convenient and highly sensitive detection of viral spread, and enzymatic activity correlated with the number of infectious particles released from infected cells. Furthermore, the Gaussia luciferase encoding virus allowed the sensitive quantification of the antiviral activity of the neuraminidase inhibitor (NAI) zanamivir and the host cell interferon-inducible transmembrane (IFITM) proteins 1-3, which are known to inhibit influenza virus entry. Finally, the virus was used to demonstrate that influenza A virus infection is sensitive to a modulator of endosomal cholesterol, in keeping with the concept that IFITMs inhibit viral entry by altering cholesterol levels in the endosomal membrane. In sum, we report the characterization of a novel influenza A reporter virus, which allows fast and sensitive detection of viral spread and its inhibition, and we show that influenza A virus entry is sensitive to alterations of endosomal cholesterol levels.
format	article
author	Nadine Eckert Florian Wrensch Sabine Gärtner Navaneethan Palanisamy Ulrike Goedecke Nils Jäger Stefan Pöhlmann Michael Winkler
author_facet	Nadine Eckert Florian Wrensch Sabine Gärtner Navaneethan Palanisamy Ulrike Goedecke Nils Jäger Stefan Pöhlmann Michael Winkler
author_sort	Nadine Eckert
title	Influenza A virus encoding secreted Gaussia luciferase as useful tool to analyze viral replication and its inhibition by antiviral compounds and cellular proteins.
title_short	Influenza A virus encoding secreted Gaussia luciferase as useful tool to analyze viral replication and its inhibition by antiviral compounds and cellular proteins.
title_full	Influenza A virus encoding secreted Gaussia luciferase as useful tool to analyze viral replication and its inhibition by antiviral compounds and cellular proteins.
title_fullStr	Influenza A virus encoding secreted Gaussia luciferase as useful tool to analyze viral replication and its inhibition by antiviral compounds and cellular proteins.
title_full_unstemmed	Influenza A virus encoding secreted Gaussia luciferase as useful tool to analyze viral replication and its inhibition by antiviral compounds and cellular proteins.
title_sort	influenza a virus encoding secreted gaussia luciferase as useful tool to analyze viral replication and its inhibition by antiviral compounds and cellular proteins.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/1c94ba363cd24da1a82b21e43ebe0ccc
work_keys_str_mv	AT nadineeckert influenzaavirusencodingsecretedgaussialuciferaseasusefultooltoanalyzeviralreplicationanditsinhibitionbyantiviralcompoundsandcellularproteins AT florianwrensch influenzaavirusencodingsecretedgaussialuciferaseasusefultooltoanalyzeviralreplicationanditsinhibitionbyantiviralcompoundsandcellularproteins AT sabinegartner influenzaavirusencodingsecretedgaussialuciferaseasusefultooltoanalyzeviralreplicationanditsinhibitionbyantiviralcompoundsandcellularproteins AT navaneethanpalanisamy influenzaavirusencodingsecretedgaussialuciferaseasusefultooltoanalyzeviralreplicationanditsinhibitionbyantiviralcompoundsandcellularproteins AT ulrikegoedecke influenzaavirusencodingsecretedgaussialuciferaseasusefultooltoanalyzeviralreplicationanditsinhibitionbyantiviralcompoundsandcellularproteins AT nilsjager influenzaavirusencodingsecretedgaussialuciferaseasusefultooltoanalyzeviralreplicationanditsinhibitionbyantiviralcompoundsandcellularproteins AT stefanpohlmann influenzaavirusencodingsecretedgaussialuciferaseasusefultooltoanalyzeviralreplicationanditsinhibitionbyantiviralcompoundsandcellularproteins AT michaelwinkler influenzaavirusencodingsecretedgaussialuciferaseasusefultooltoanalyzeviralreplicationanditsinhibitionbyantiviralcompoundsandcellularproteins
_version_	1718421910236168192

Influenza A virus encoding secreted Gaussia luciferase as useful tool to analyze viral replication and its inhibition by antiviral compounds and cellular proteins.

Ejemplares similares