Biophysical Characterization of the Complex between Double-Stranded RNA and the N-Terminal Domain of the NS1 Protein from Influenza A Virus:  Evidence for a Novel RNA-Binding Mode†

Biochemistry ◽  
2004 ◽  
Vol 43 (7) ◽  
pp. 1950-1962 ◽  
Author(s):  
Chen-ya Chien ◽  
Yujia Xu ◽  
Rong Xiao ◽  
James M. Aramini ◽  
Parag V. Sahasrabudhe ◽  
...  
Keyword(s):  
Influenza A Virus ◽  
Influenza A ◽  
Rna Binding ◽  
Binding Mode ◽  
Ns1 Protein ◽  
Biophysical Characterization ◽  
Double Stranded Rna ◽  
Terminal Domain

scholarly journals Binding of the influenza A virus NS1 protein to PKR mediates the inhibition of its activation by either PACT or double-stranded RNA

Virology ◽  
2006 ◽  
Vol 349 (1) ◽  
pp. 13-21 ◽  
Author(s):  
Shoudong Li ◽  
Ji-Young Min ◽  
Robert M. Krug ◽  
Ganes C. Sen
Keyword(s):  
Influenza A Virus ◽  
Influenza A ◽  
Ns1 Protein ◽  
Double Stranded Rna

scholarly journals The accumulation of influenza A virus segment 7 spliced mRNAs is regulated by the NS1 protein

2012 ◽  
Vol 93 (1) ◽  
pp. 113-118 ◽  
Author(s):  
Nicole C. Robb ◽  
Ervin Fodor
Keyword(s):  
Influenza Virus ◽  
Viral Infection ◽  
Influenza A Virus ◽  
Influenza A ◽  
Rna Binding ◽  
Mrna Splicing ◽  
Ns1 Protein ◽  
Binding Ability ◽  
Alternatively Spliced ◽  
Transcription And Replication

The influenza A virus M1 mRNA is alternatively spliced to produce M2 mRNA, mRNA3, and in some cases, M4 mRNA. Splicing of influenza mRNAs is carried out by the cellular splicing machinery and is thought to be regulated, as both spliced and unspliced mRNAs encode proteins. In this study, we used radioactively labelled primers to investigate the accumulation of spliced and unspliced M segment mRNAs in viral infection and ribonucleoprotein (RNP) reconstitution assays in which only the minimal components required for transcription and replication to occur were expressed. We found that co-expression of the viral NS1 protein in an RNP reconstitution assay altered the accumulation of spliced mRNAs compared with when it was absent, and that this activity was dependent on the RNA-binding ability of NS1. These findings suggest that the NS1 protein plays a role in the regulation of splicing of influenza virus M1 mRNA.

scholarly journals Characterization of a Novel Mutation in NS1 Protein of Influenza A Virus Induced by a Chemical Substance for the Attenuation of Pathogenicity

PLoS ONE ◽  
2015 ◽  
Vol 10 (3) ◽  
pp. e0121205 ◽  
Author(s):  
Kohei Sasaki ◽  
Kyoko Hayashi ◽  
Jung-Bum Lee ◽  
Fumiya Kurosaki ◽  
Toshimitsu Hayashi
Keyword(s):  
Influenza A Virus ◽  
Influenza A ◽  
Novel Mutation ◽  
Chemical Substance ◽  
Ns1 Protein

scholarly journals Functional Replacement of the Carboxy-Terminal Two-Thirds of the Influenza A Virus NS1 Protein with Short Heterologous Dimerization Domains

2002 ◽  
Vol 76 (24) ◽  
pp. 12951-12962 ◽  
Author(s):  
Xiuyan Wang ◽  
Christopher F. Basler ◽  
Bryan R. G. Williams ◽  
Robert H. Silverman ◽  
Peter Palese ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Influenza A Virus ◽  
Influenza A ◽  
Rna Binding ◽  
Influenza Viruses ◽  
Ns1 Protein ◽  
Wild Type ◽  
Dimerization Domain ◽  
Carboxy Terminal

ABSTRACT The NS1 protein of influenza A/WSN/33 virus is a 230-amino-acid-long protein which functions as an interferon alpha/beta (IFN-α/β) antagonist by preventing the synthesis of IFN during viral infection. In tissue culture, the IFN inhibitory function of the NS1 protein has been mapped to the RNA binding domain, the first 73 amino acids. Nevertheless, influenza viruses expressing carboxy-terminally truncated NS1 proteins are attenuated in mice. Dimerization of the NS1 protein has previously been shown to be essential for its RNA binding activity. We have explored the ability of heterologous dimerization domains to functionally substitute in vivo for the carboxy-terminal domains of the NS1 protein. Recombinant influenza viruses were generated that expressed truncated NS1 proteins of 126 amino acids, fused to 28 or 24 amino acids derived from the dimerization domains of either the Saccharomyces cerevisiae PUT3 or the Drosophila melanogaster Ncd (DmNcd) proteins. These viruses regained virulence and lethality in mice. Moreover, a recombinant influenza virus expressing only the first 73 amino acids of the NS1 protein was able to replicate in mice lacking three IFN-regulated antiviral enzymes, PKR, RNaseL, and Mx, but not in wild-type (Mx-deficient) mice, suggesting that the attenuation was mainly due to an inability to inhibit the IFN system. Remarkably, a virus with an NS1 truncated at amino acid 73 but fused to the dimerization domain of DmNcd replicated and was also highly pathogenic in wild-type mice. These results suggest that the main biological function of the carboxy-terminal region of the NS1 protein of influenza A virus is the enhancement of its IFN antagonist properties by stabilizing the NS1 dimeric structure.

Antiviral activity of double‐stranded RNA‐binding protein PACT against influenza A virus mediated via suppression of viral RNA polymerase

2018 ◽  
Vol 32 (8) ◽  
pp. 4380-4393 ◽  
Author(s):  
Chi‐Ping Chan ◽  
Chun‐Kit Yuen ◽  
Pak‐Hin Hinson Cheung ◽  
Sin‐Yee Fung ◽  
Pak‐Yin Lui ◽  
...  
Keyword(s):  
Antiviral Activity ◽  
Rna Polymerase ◽  
Influenza A Virus ◽  
Influenza A ◽  
Rna Binding ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
Viral Rna ◽  
Double Stranded Rna

The N-terminal residual arginine19 of influenza A virus NS1 protein is required for its nuclear localization and RNA binding

2020 ◽  
Vol 251 ◽  
pp. 108895
Author(s):  
Xingbo Wang ◽  
Lulu Lin ◽  
Yang Yu ◽  
Yan Yan ◽  
Nishant Kumar Ojha ◽  
...  
Keyword(s):  
Influenza A Virus ◽  
Nuclear Localization ◽  
Influenza A ◽  
Rna Binding ◽  
Ns1 Protein

scholarly journals The Influenza A Virus NS1 Protein Inhibits Activation of Jun N-Terminal Kinase and AP-1 Transcription Factors

2002 ◽  
Vol 76 (21) ◽  
pp. 11166-11171 ◽  
Author(s):  
Stephan Ludwig ◽  
Xiuyan Wang ◽  
Christina Ehrhardt ◽  
Hongyong Zheng ◽  
Nicola Donelan ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcription Factors ◽  
Stress Response ◽  
Influenza A Virus ◽  
Influenza A ◽  
Ns1 Protein ◽  
Double Stranded Rna ◽  
Antiviral Responses ◽  
Cell Gene Expression ◽  
Cell Gene

ABSTRACT The influenza A virus nonstructural NS1 protein is known to modulate host cell gene expression and to inhibit double-stranded RNA (dsRNA)-mediated antiviral responses. Here we identify NS1 as the first viral protein that antagonizes virus- and dsRNA-induced activation of the stress response-signaling pathway mediated through Jun N-terminal kinase.

scholarly journals Robust Lys63-Linked Ubiquitination of RIG-I Promotes Cytokine Eruption in Early Influenza B Virus Infection

2016 ◽  
Vol 90 (14) ◽  
pp. 6263-6275 ◽  
Author(s):  
Jingwen Jiang ◽  
Jing Li ◽  
Wenhui Fan ◽  
Weinan Zheng ◽  
Meng Yu ◽  
...  
Keyword(s):  
Virus Infection ◽  
Influenza A Virus ◽  
Influenza A ◽  
Molecular Mechanisms ◽  
Rna Binding ◽  
Influenza B Virus ◽  
Influenza B ◽  
Type I ◽  
Ns1 Protein ◽  
B Virus

ABSTRACTInfluenza A and B virus infections both cause a host innate immunity response. Here, we report that the robust production of type I and III interferons (IFNs), IFN-stimulated genes, and proinflammatory factors can be induced by influenza B virus rather than influenza A virus infection in alveolar epithelial (A549) cells during early infection. This response is mainly dependent on the retinoic acid-inducible gene I (RIG-I)-mediated signaling pathway. Infection by influenza B virus promotes intense Lys63-linked ubiquitination of RIG-I, resulting in cytokine eruption. It is known that the influenza A virus NS1 protein (NS1-A) interacts with RIG-I and TRIM25 to suppress the activation of RIG-I-mediated signaling. However, the present results indicate that the influenza B virus NS1 protein (NS1-B) is unable to interact with RIG-I but engages in the formation of a RIG-I/TRIM25/NS1-B ternary complex. Furthermore, we demonstrate that the N-terminal RNA-binding domain (RBD) of NS1-B is responsible for interaction with TRIM25 and that this interaction blocks the inhibitory effect of the NS1-B C-terminal effector domain (TED) on RIG-I ubiquitination. Our findings reveal a novel mechanism for the host cytokine response to influenza B virus infection through regulatory interplay between host and viral proteins.IMPORTANCEInfluenza B virus generally causes local mild epidemics but is occasionally lethal to individuals. Existing studies describe the broad characteristics of influenza B virus epidemiology and pathology. However, to develop better prevention and treatments for the disease, determining the concrete molecular mechanisms of pathogenesis becomes pivotal to understand how the host reacts to the challenge of influenza B virus. Thus, we aimed to characterize the host innate immune response to influenza B virus infection. Here, we show that vigorous Lys63-linked ubiquitination of RIG-I and cytokine eruption dependent on RIG-I-mediated signal transduction are induced by virus infection. Additionally, TRIM25 positively regulates RIG-I-mediated signaling by ablating the inhibitory function of NS1-B on RIG-I ubiquitination.

scholarly journals ISG15 conjugation system targets the viral NS1 protein in influenza A virus–infected cells

2010 ◽  
Vol 107 (5) ◽  
pp. 2253-2258 ◽  
Author(s):  
Chen Zhao ◽  
Tien-Ying Hsiang ◽  
Rei-Lin Kuo ◽  
Robert M. Krug
Keyword(s):  
Antiviral Activity ◽  
Influenza A Virus ◽  
Viral Protein ◽  
Influenza A ◽  
Rna Binding ◽  
Acceptor Site ◽  
Ns1 Protein ◽  
Conjugation System ◽  
Infected Cells ◽  
Ns1a Protein

ISG15 is an IFN-α/β–induced, ubiquitin-like protein that is conjugated to a wide array of cellular proteins through the sequential action of three conjugation enzymes that are also induced by IFN-α/β. Recent studies showed that ISG15 and/or its conjugates play an important role in protecting cells from infection by several viruses, including influenza A virus. However, the mechanism by which ISG15 modification exerts antiviral activity has not been established. Here we extend the repertoire of ISG15 targets to a viral protein by demonstrating that the NS1 protein of influenza A virus (NS1A protein), an essential, multifunctional protein, is ISG15 modified in virus-infected cells. We demonstrate that the major ISG15 acceptor site in the NS1A protein in infected cells is a critical lysine residue (K41) in the N-terminal RNA-binding domain (RBD). ISG15 modification of K41 disrupts the association of the NS1A RBD domain with importin-α, the protein that mediates nuclear import of the NS1A protein, whereas the RBD retains its double-stranded RNA-binding activity. Most significantly, we show that ISG15 modification of K41 inhibits influenza A virus replication and thus contributes to the antiviral action of IFN-β. We also show that the NS1A protein directly and specifically binds to Herc5, the major E3 ligase for ISG15 conjugation in human cells. These results establish a “loss of function” mechanism for the antiviral activity of the IFN-induced ISG15 conjugation system, namely, that it inhibits viral replication by conjugating ISG15 to a specific viral protein, thereby inhibiting its function.

Sign in / Sign up

Export Citation Format

Share Document