Nuclear dynamics of influenza A virus ribonucleoproteins revealed by live-cell imaging studies.

The negative sense RNA genome of influenza A virus is transcribed and replicated in the nuclei of infected cells by the viral RNA polymerase. Only four viral polypeptides are required but multiple cellular components are potentially involved. We used fluorescence recovery after photobleaching (FRAP)...

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Main Authors: Loucaides, E, von Kirchbach, J, Foeglein, A, Sharps, J, Fodor, E, Digard, P
Format: Journal article
Language:English
Published: 2009
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author Loucaides, E
von Kirchbach, J
Foeglein, A
Sharps, J
Fodor, E
Digard, P
author_facet Loucaides, E
von Kirchbach, J
Foeglein, A
Sharps, J
Fodor, E
Digard, P
author_sort Loucaides, E
collection OXFORD
description The negative sense RNA genome of influenza A virus is transcribed and replicated in the nuclei of infected cells by the viral RNA polymerase. Only four viral polypeptides are required but multiple cellular components are potentially involved. We used fluorescence recovery after photobleaching (FRAP) to characterise the dynamics of GFP-tagged viral ribonucleoprotein (RNP) components in living cells. The nucleoprotein (NP) displayed very slow mobility that significantly increased on formation of transcriptionally active RNPs. Conversely, single or dimeric polymerase subunits showed fast nuclear dynamics that decreased upon formation of heterotrimers, suggesting increased interaction of the full polymerase complex with a relatively immobile cellular component(s). Treatment with inhibitors of cellular transcription indicated that in part, this reflected an interaction with cellular RNA polymerase II. Analysis of mutated influenza virus polymerase complexes further suggested that this was through an interaction between PB2 and RNA Pol II separate from PB2 cap-binding activity.
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spelling oxford-uuid:bb3f9513-664f-42c1-899e-858addc847962022-03-27T05:15:40ZNuclear dynamics of influenza A virus ribonucleoproteins revealed by live-cell imaging studies.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:bb3f9513-664f-42c1-899e-858addc84796EnglishSymplectic Elements at Oxford2009Loucaides, Evon Kirchbach, JFoeglein, ASharps, JFodor, EDigard, PThe negative sense RNA genome of influenza A virus is transcribed and replicated in the nuclei of infected cells by the viral RNA polymerase. Only four viral polypeptides are required but multiple cellular components are potentially involved. We used fluorescence recovery after photobleaching (FRAP) to characterise the dynamics of GFP-tagged viral ribonucleoprotein (RNP) components in living cells. The nucleoprotein (NP) displayed very slow mobility that significantly increased on formation of transcriptionally active RNPs. Conversely, single or dimeric polymerase subunits showed fast nuclear dynamics that decreased upon formation of heterotrimers, suggesting increased interaction of the full polymerase complex with a relatively immobile cellular component(s). Treatment with inhibitors of cellular transcription indicated that in part, this reflected an interaction with cellular RNA polymerase II. Analysis of mutated influenza virus polymerase complexes further suggested that this was through an interaction between PB2 and RNA Pol II separate from PB2 cap-binding activity.
spellingShingle Loucaides, E
von Kirchbach, J
Foeglein, A
Sharps, J
Fodor, E
Digard, P
Nuclear dynamics of influenza A virus ribonucleoproteins revealed by live-cell imaging studies.
title Nuclear dynamics of influenza A virus ribonucleoproteins revealed by live-cell imaging studies.
title_full Nuclear dynamics of influenza A virus ribonucleoproteins revealed by live-cell imaging studies.
title_fullStr Nuclear dynamics of influenza A virus ribonucleoproteins revealed by live-cell imaging studies.
title_full_unstemmed Nuclear dynamics of influenza A virus ribonucleoproteins revealed by live-cell imaging studies.
title_short Nuclear dynamics of influenza A virus ribonucleoproteins revealed by live-cell imaging studies.
title_sort nuclear dynamics of influenza a virus ribonucleoproteins revealed by live cell imaging studies
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