Influenza A virus nucleoprotein, is a multifunctional RNA-binding protein, encoded by

Influenza A virus nucleoprotein, is a multifunctional RNA-binding protein, encoded by segment-5 of the negative sense RNA genome. cells. Depletion of nucleolin in A549 buy 1370554-01-0 cells by siRNA targeting endogenous nucleolin followed by influenza A virus contamination, disrupted its conversation with viral nucleoprotein, resulting in increased expression of gene transcripts encoding late viral protein; matrix (M1) and hemagglutinin (HA) in infected cells. On the contrary, over expression of nucleolin in cells transiently transfected with pEGFP-NCL construct followed by virus contamination significantly reduced the late viral gene transcripts, and consequently the viral titer. Altered expression of late viral genes and titers following manipulation of host cellular nucleolin, proposes the functional importance of its conversation with nucleoprotein during influenza A virus contamination. Introduction Influenza A virus is usually a public health threat worldwide and contributes to a high-level of mortality during pandemics. Its segmented genetic composition allows re-assortment of gene segments between the strains of different host origins resulting in the emergence of a novel strain and an unpredictable pandemic. Eight unfavorable sense single stranded RNA gene segments of influenza A virus encode for 10 proteins. However, 7 more novel proteins; PB1-F2 [1], PB1-N40 [2], PA-N155, PA-N182 [3], PA-X [4], M42 [5] and NS3 [6] were discovered 40years after the genome mapping of influenza A virus was done. Nucleoprotein (NP), a 56kDa protein, encoded by segment-5, is usually a multifunctional RNA binding protein. The primary function of this protein is usually to encapsidate the viral genome and form homo-oligomers to maintain ribonucleoprotein complex (RNP) structure [7]. It acts as a cofactor to coat the newly synthesized viral complementary RNA [8]. Also, NP determines whether complementary RNA, synthesized from genomic RNA, is usually to be used for protein translation or to serve as a template for synthesis of genomic strand during the replication [9]. Besides its active role in the replication buy 1370554-01-0 of virus, NP contributes to host adaptation when avian strains change their host to mammalian species. During adaptation, mutations in NP together with subunits of RNA polymerase contribute to increased polymerase activity of avian strains [10]. All the above reports together signify the indispensable role of NP in influenza A virus life cycle. Owing to their limited genetic coding capacity, viruses depend on the host machinery for their survival. The virus and the host protein interactions in infected cell during virus replication determine the outcome of the disease. Conversation of host protein with influenza A virus protein have been reported earlier in the literature using various approaches such as genome-wide RNA interference screening [11], yeast-two-hybrid screening [12], comprehensive analysis of influenza virus polymerase cellular interactome [13] and proteome based approaches [14]. As influenza virus replicates Pax6 inside the nucleus, NP constantly shuttles between the nucleus and the cytoplasm interacting with various host factors to regulate multiple functions during the replication [15]. Viral RNP complex is usually imported into the nucleus through its conversation with importin [16], and viral RNA synthesis is usually regulated by conversation between NP and cellular factors such as BAT1/UAP56, Tat-SF1, MCM complexes [17, 18, 19]. So is usually export of vRNA into the cytoplasm mediated by NP conversation with CRM1 [20]. Furthermore, NP association with cytoskeletal proteins buy 1370554-01-0 such as alpha actinin-4 exhibited to be essential for its translocation into the nucleus [21]. Nonetheless, the host interacting partners to NP of 2009 pandemic H1N1 strain and the interacting partners that are common to NP of circulating and emerging strains of influenza A virus are largely unknown. Since, the gene encoding viral NP is usually conserved among the strains isolated from different hosts with less than 11% of the amino acid difference [22], NP of different influenza A viral strains are expected to have common host interacting partners. In the current study, we sought to identify the novel host interacting partners.