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Título: | The host-dependent interaction of alpha-importins with influenza PB2 polymerase subunit is required for virus RNA replication |
Autor: | Resa-Infante, Patricia; Jorba, Núria; Zamarreño, Noelia; Fernández, Yolanda; Juárez, Silvia; Ortín, Juan CSIC ORCID | Fecha de publicación: | 10-dic-2008 | Editor: | Public Library of Science | Citación: | PLoS ONE 3(12): e3904 (2008) | Resumen: | The influenza virus polymerase is formed by the PB1, PB2 and PA subunits and is required for virus transcription and replication in the nucleus of infected cells. As PB2 is a relevant host-range determinant we expressed a TAP-tagged PB2 in human cells and isolated intracellular complexes. Alpha-importin was identified as a PB2-associated factor by proteomic analyses. To study the relevance of this interaction for virus replication we mutated the PB2 NLS and analysed the phenotype of mutant subunits, polymerase complexes and RNPs. While mutant PB2 proteins showed reduced nuclear accumulation, they formed polymerase complexes normally when co expressed with PB1 and PA. However, mutant RNPs generated with a viral CAT replicon showed up to hundred-fold reduced CAT accumulation. Rescue of nuclear localisation of mutant PB2 by insertion of an additional SV40 TAg-derived NLS did not revert the mutant phenotype of RNPs. Furthermore, determination of recombinant RNP accumulation in vivo indicated that PB2 NLS mutations drastically reduced virus RNA replication. These results indicate that, above and beyond its role in nuclear accumulation, PB2 interaction with α-importins is required for virus RNA replication. To ascertain whether PB2-α-importin binding could contribute to the adaptation of H5N1 avian viruses to man, their association in vivo was determined. Human alpha importin isoforms associated efficiently to PB2 protein of an H3N2 human virus but bound to diminished and variable extents to PB2 from H5N1 avian or human strains, suggesting that the function of alpha importin during RNA replication is important for the adaptation of avian viruses to the human host. | Descripción: | 10 pages, 8 figures.-- PMID: 19066626 [PubMed].-- PMCID: PMC2588535.-- Supporting information (Suppl. figures S1-S2 & tables S1-S2) available at: http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0003904 | Versión del editor: | http://dx.doi.org/10.1371/journal.pone.0003904 | URI: | http://hdl.handle.net/10261/11878 | DOI: | 10.1371/journal.pone.0003904 | ISSN: | 1932-6203 |
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Fichero | Descripción | Tamaño | Formato | |
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Resa_et_al_PLoS_ONE_3_12_2008.pdf | Main text file | 1,27 MB | Adobe PDF | Visualizar/Abrir |
journal.pone.0003904.s001.tiff | Fig. S1 | 1,62 MB | TIFF | Visualizar/Abrir |
journal.pone.0003904.s002.tiff | Fig. S2 | 5,27 MB | TIFF | Visualizar/Abrir |
journal.pone.0003904.s003.doc | Table S1 | 27,5 kB | Microsoft Word | Visualizar/Abrir |
journal.pone.0003904.s004.doc | Table S2 | 128,5 kB | Microsoft Word | Visualizar/Abrir |
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