tetano
Editor, Senior Moderator
FEBS J. 2014 May 7. doi: 10.1111/febs.12828. [Epub ahead of print]
Influenza A Viral Nucleoprotein Interacts with Cytoskeleton Scaffolding Protein α-actinin-4 for Viral Replication.
Sharma S1, Mayank AK, Nailwal H, Tripathi S, Patel JR, Bradford Bowzard J, Gaur P, Donis RO, Katz JM, Cox NJ, Lal RB, Narula A, Sambhara S, Lal SK.
Author information
Abstract
Influenza A virus (IAV), like other viruses, exploits the machinery of human host cells for its survival and replication. We identified α-actinin-4, a host cytoskeletal protein, as an interacting partner of IAV nucleoprotein (NP). We confirmed this interaction using co-immunoprecipitation studies first in coupled in-vitro transcription-translation assay and then in cells either transiently co-expressing the two proteins or infected with whole IAV. Importantly, the NP-actinin-4 interaction was observed in several IAV subtypes including the 2009 H1N1 pandemic virus. Moreover, immunofluorescence studies revealed that both NP and actinin-4 co-localized largely around the nucleus and also in the cytoplasmic region of virus-infected A549 cells. Silencing of actinin-4 expression resulted in not only a significant decrease in NP, M2 and NS1 viral protein expression but also reduction of both, NP mRNA and vRNA levels as well as viral titers, 24 hr post-infection with Influenza A virus, suggesting that actinin-4 was critical for viral replication. Furthermore, actinin-4 depletion reduced the amount of NP localized in the nucleus. Treatment of infected cells with wortmannin, a known inhibitor of actinin-4, led to decrease in NP mRNA levels and also caused nuclear retention of NP, further strengthening our previous observations. Taken together our results indicate that actinin-4, a novel interacting partner of IAV NP, plays a crucial role in viral replication and this interaction may participate in nuclear localization of NP and/or vRNPs This article is protected by copyright. All rights reserved.
STRUCTURED DIGITAL ABSTRACT:
NP physically interacts with actinin-4 by anti bait coimmunoprecipitation (1,2) NP and actnin-4 colocalize by fluorescence microscopy (View interaction) NP physically interacts with actinin-4 by anti bait coimmunoprecipitation (View interaction) NP binds to actinin-4 by anti tag coimmunoprecipitation (1, 2) NP physically interacts with actinin-4 by anti bait coimmunoprecipitation (View interaction) NP physically interacts with actinin-4 by anti tag coimmunoprecipitation (View interaction) NP physically interacts with actinin-4 by anti bait coimmunoprecipitation (View interaction) NP physically interacts with actinin-4 by anti bait coimmunoprecipitation (View interaction) NP physically interacts with actinin-4 by two hybrid (1,2) NP physically interacts with actinin-4 by anti bait coimmunoprecipitation (View interaction).
This article is protected by copyright. All rights reserved.
KEYWORDS:
Actinin-4, Influenza virus, Interaction, Nucleoprotein, localization, virus titer
PMID:
24802111
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24802111
Influenza A Viral Nucleoprotein Interacts with Cytoskeleton Scaffolding Protein α-actinin-4 for Viral Replication.
Sharma S1, Mayank AK, Nailwal H, Tripathi S, Patel JR, Bradford Bowzard J, Gaur P, Donis RO, Katz JM, Cox NJ, Lal RB, Narula A, Sambhara S, Lal SK.
Author information
Abstract
Influenza A virus (IAV), like other viruses, exploits the machinery of human host cells for its survival and replication. We identified α-actinin-4, a host cytoskeletal protein, as an interacting partner of IAV nucleoprotein (NP). We confirmed this interaction using co-immunoprecipitation studies first in coupled in-vitro transcription-translation assay and then in cells either transiently co-expressing the two proteins or infected with whole IAV. Importantly, the NP-actinin-4 interaction was observed in several IAV subtypes including the 2009 H1N1 pandemic virus. Moreover, immunofluorescence studies revealed that both NP and actinin-4 co-localized largely around the nucleus and also in the cytoplasmic region of virus-infected A549 cells. Silencing of actinin-4 expression resulted in not only a significant decrease in NP, M2 and NS1 viral protein expression but also reduction of both, NP mRNA and vRNA levels as well as viral titers, 24 hr post-infection with Influenza A virus, suggesting that actinin-4 was critical for viral replication. Furthermore, actinin-4 depletion reduced the amount of NP localized in the nucleus. Treatment of infected cells with wortmannin, a known inhibitor of actinin-4, led to decrease in NP mRNA levels and also caused nuclear retention of NP, further strengthening our previous observations. Taken together our results indicate that actinin-4, a novel interacting partner of IAV NP, plays a crucial role in viral replication and this interaction may participate in nuclear localization of NP and/or vRNPs This article is protected by copyright. All rights reserved.
STRUCTURED DIGITAL ABSTRACT:
NP physically interacts with actinin-4 by anti bait coimmunoprecipitation (1,2) NP and actnin-4 colocalize by fluorescence microscopy (View interaction) NP physically interacts with actinin-4 by anti bait coimmunoprecipitation (View interaction) NP binds to actinin-4 by anti tag coimmunoprecipitation (1, 2) NP physically interacts with actinin-4 by anti bait coimmunoprecipitation (View interaction) NP physically interacts with actinin-4 by anti tag coimmunoprecipitation (View interaction) NP physically interacts with actinin-4 by anti bait coimmunoprecipitation (View interaction) NP physically interacts with actinin-4 by anti bait coimmunoprecipitation (View interaction) NP physically interacts with actinin-4 by two hybrid (1,2) NP physically interacts with actinin-4 by anti bait coimmunoprecipitation (View interaction).
This article is protected by copyright. All rights reserved.
KEYWORDS:
Actinin-4, Influenza virus, Interaction, Nucleoprotein, localization, virus titer
PMID:
24802111
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/24802111